Isoprenyl Compounds and Methods Thereof

ABSTRACT

Among other things, the present invention provides novel isoprenyl compounds capable of effectively modulating inflammatory responses and pharmaceutical, cosmetic, cosmeceutical and topical compositions comprising these isoprenyl compounds. Anti-inflammatory compounds of the present invention are useful in treating or preventing diseases or conditions associated with inflammation. Proinflammatory compounds of the present invention are useful in treating or preventing diseases or conditions associated with suppression of inflammatory responses. Thus, the present invention also provides methods useful in the treatment or prevention of diseases or conditions associated with inflammation as well as methods useful in the treatment or prevention of diseases or conditions associated with suppression of inflammatory responses.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.13/760,627 filed on Feb. 6, 2013, now U.S. Pat. No. 9,376,383, which isa continuation of U.S. patent application Ser. No. 12/616,781 filed onNov. 12, 2009, now U.S. Pat. No. 8,372,884, which claims priority fromU.S. Provisional Patent Application No. 61/113,498 filed on Nov. 11,2008, the entire disclosures of each of the aforementioned applicationsare incorporated herein by reference.

GOVERNMENT LICENSE RIGHTS

This invention was made with government support under SBIR grants1R43AI062034-01A2 and R44AI062034 awarded by National Institutes ofHealth. The government has certain rights in the invention.

BACKGROUND

Inflammation often is a bodily response to infection or injury in whichcells involved in detoxification and repair are mobilized to thecompromised site by inflammatory mediators. Such infection or injury canbe a result of acute or chronic disease, disorders, conditions ortrauma, or of environmental conditions or aging. Examples includediseases, disorders, syndromes, conditions and injuries of thecardiovascular, digestive, integumentary, muscular, nervous,reproductive, respiratory and urinary systems, as well as, diseases,disorders, syndromes, conditions and injuries of tissue and cartilagesuch as atherosclerosis, irritable bowel syndrome, psoriasis,tendonitis, Alzheimer's disease and vascular dementia, multiplesclerosis, diabetes, endometriosis, asthma and kidney failure. Treatmentof inflammatory diseases or disorders with traditional anti-inflammatorydrugs, e.g., corticosteroids and non-steroidal anti-inflammatory drugs(“NSAIDS”) can cause multiple side effects, e.g., appetite and weightgain, excess sweating, high blood pressure, nausea, vomiting, diarrhea,etc.

SUMMARY

The present invention provides, among other things, certain compoundsthat are structurally related to N-acetyl-S-farnesyl-L-cysteine (“AFC”).

In some embodiments, the present invention provides a compound ofFormula I:

or a pharmaceutically acceptable salt thereof, wherein:

L is a bivalent, branched or unbranched, saturated or unsaturated, C₂-C₆hydrocarbon chain wherein one or more methylene units of L isindependently replaced by —O—, —S—, —NH—, —C(O)—, —C(═CH₂)—, or C₃-C₆cycloalkylene, wherein L is optionally substituted by one or more groupsselected from halogen, phenyl, an 8-10 membered bicyclic aryl ring, a5-6 membered heteroaryl ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, an 8-10 membered bicyclicheteroaryl ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, a 5- to 7-membered monocyclic having 1-2heteroatoms independently selected from nitrogen, oxygen, or sulfur or a7-10 membered bicyclic heterocyclyl ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, or sulfur;

R¹ is hydrogen, —OH or —OR, wherein each R is independently hydrogen oran optionally substituted group selected from C₁₋₆ aliphatic or C₁₋₆heteroaliphatic;

R² is —C(O)X, wherein X is independently R, —OR, a hydrogen, aryloxy,amino, alkylamino, dialkylamino, heteroaryloxy, hydrazine, a 6-10membered aryl ring, a 5-6 membered heteroaryl ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein each R is independently hydrogen or an optionally substitutedgroup selected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic; and

R³ is a substituted or unsubstituted, branched or unbranched, saturatedor unsaturated, C₁₀-C₂₅ aliphatic.

In some embodiments, the present invention provides a compound ofFormula I′:

or a pharmaceutically acceptable salt thereof, wherein:

L is a bivalent, branched or unbranched, saturated or unsaturated, C₂-C₆hydrocarbon chain wherein one or more methylene units of L isindependently replaced by —O—, —S—, —NH—, —C(O)—, —CF₂—, —C(═CH₂)—,—CH═CH—, or an optionally substituted arylene, heteroarylene, C₃-C₆cycloalkylene, C₃-C₆ heterocycloalkylene, or an 8-10-membered bicyclicheterocyclic moiety,

and wherein L is optionally substituted by one or more groups selectedfrom halogen, C₁-C₆ alkyl, phenyl, biphenyl, -benzyl, —CH₂-phenol,—CH(phenyl)₂, —OH, —NH₂, —NHC(O)CH₃, —NHC(O)NHCH₂CH₃, —C(O)NH₂,—C(O)NHCH₂CH₃, —CH₂C(O)OCH₂phenyl, —(CH₂)₂SCH₃, —(CH₂)₂C(O)NH₂,—(CH₂)₂C(O)OH, an 8-10 membered bicyclic aryl ring, a 5-6 memberedheteroaryl ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, an 8-10 membered bicyclic heteroaryl ringhaving 1-4 heteroatoms independently selected from nitrogen, oxygen, orsulfur, a 5- to 7-membered monocyclic having 1-2 heteroatomsindependently selected from nitrogen, oxygen, or sulfur or a 7-10membered bicyclic heterocyclyl ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur;

M is —C(O)—, —C(S), or —SO₂—;

R¹ is hydrogen, F, CF₃, C₁-C₄ alkyl, —OH, —C(O)CH₃, —NH(OR), —NR₂,—NHNR₂, —SO₂R, —NH-phenyl, —SO₂-phenyl, -phenyl-NO₂, or —OR, whereineach R is independently hydrogen, oxygen, or an optionally substitutedgroup selected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic;

R² is —C(O)X, wherein X is independently R, —C(O)NHNH₂, —OR, a hydrogen,aryloxy, amino, alkylamino, dialkylamino, heteroaryloxy, hydrazine, a6-10 membered aryl ring, a 5-6 membered heteroaryl ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein each R is independently hydrogen or an optionally substitutedgroup selected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic; and

R³ is a substituted or unsubstituted, branched or unbranched, saturatedor unsaturated, C₁₀-C₂₅ aliphatic; and

Y is —O—, —N—, —S—, —Se—, —S(O)—, —S(═N)—, —SO₂—, —Se(O)—, or —Se(O)₂—.

In some embodiments, compounds of Formulae I and/or I′ are provided withthe proviso that L and R¹ cannot together be C₁-C₃ unsubstitutednon-halogenated alkyl.

In certain embodiments, the present invention provides a compound ofFormula Ia,

or a pharmaceutically acceptable salt thereof, wherein:

L is a bivalent, branched or unbranched, saturated or unsaturated, C₂-C₆hydrocarbon chain wherein one or more methylene units of L isindependently replaced by —O—, —S—, —NH—, —C(O)—, —C(═CH₂)—, or C₃-C₆cycloalkylene, wherein L is optionally substituted by one or more groupsselected from halogen, phenyl, an 8-10 membered bicyclic aryl ring, a5-6 membered heteroaryl ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, an 8-10 membered bicyclicheteroaryl ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, a 5- to 7-membered monocyclic having 1-2heteroatoms independently selected from nitrogen, oxygen, or sulfur or a7-10 membered bicyclic heterocyclyl ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, or sulfur;

R¹ is hydrogen, —OH or —OR, wherein each R is independently hydrogen oran optionally substituted group selected from C₁₋₆ aliphatic or C₁₋₆heteroaliphatic; and

R² is —C(O)X, wherein X is independently R, —OR, a hydrogen, aryloxy,amino, alkylamino, dialkylamino, heteroaryloxy, hydrazine, a 6-10membered aryl ring, a 5-6 membered heteroaryl ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein each R is independently hydrogen or an optionally substitutedgroup selected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic.

In some embodiments, treatment of inflammatory diseases or disorders isachieved using compounds of the present invention.

In some embodiments, treatment of inflammatory diseases or disorders isachieved using provided compounds without having the side effects ofcorticosteroids or NSAIDS.

The present invention encompasses the finding that certain isoprenylcompounds have surprising and desirable characteristics as compared withother compounds and/or N-acetyl-S-farnesyl-L-cysteine (“AFC”). Forexample, the present disclosure illustrates that certain isoprenylcompounds show surprising inhibition of edema, erythema and dermalneutrophil infiltration, as measured by inhibition of MPO(myeloperoxidase), when compared to AFC.

In some embodiments, the present invention provides compounds thatmodulate the G-protein signaling cascade. In some embodiments, thepresent invention provides compounds that modulate inflammatorypathways. In certain embodiments, the inflammatory pathway is associatedwith G-protein pathways (e.g., purinergic receptor-mediated). In certainembodiments, the inflammatory pathway is associated with non G-proteinpathways (e.g., PPAR-mediated, Toll-like receptor-mediated, andTNF-alpha receptor-mediated). In certain embodiments, isoprenylcompounds of the present invention modulate the levels of inflammatorymediators (e.g., cytokines). In certain embodiments, isoprenyl compoundsof the present invention demonstrate surprising inhibition ofpro-inflammatory cytokines (e.g., IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5,IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12/IL-23 p40, IL13, IL-17,IL-18, TGF-β, IFN-γ, GM-CSF, Groα, MCP-1 and TNF-α).

In some embodiments, isoprenyl compounds of the present inventionexhibit anti-oxidant activity.

In some embodiments, isoprenyl compounds of the present inventiondemonstrate surprising inhibition of helper T-lymphocyte infiltrationand accumulation.

In some embodiments, isoprenyl compounds of the present invention showsurprising inhibition of oxidative burst response from neutrophils.

In some embodiments, compounds provided by the present invention havethe structure set forth in Formulae I, I′, and/or Ia herein. Certainparticular embodiments are described in more detail below.

The present invention also provides compositions containing compoundsdescribed herein, methods of preparing such compounds and/orcompositions, and methods of using such compounds and/or compositions.

In some embodiments, the present invention provides compositions fortreating or preventing inflammation comprising at least one isoprenylcompound, a carrier and optionally an additional active ingredient.

In some embodiments, the present invention provides topical compositionsfor treating or preventing a skin disease or condition, comprising atleast one isoprenyl compound, a carrier and optionally an additionalactive ingredient, formulated for topical administration. In certainembodiments, provided herein are topical compositions for promotinghealthy skin in a subject comprising at least one isoprenyl compound, acarrier and optionally an additional active ingredient.

In some embodiments, the present invention provides compositions fortreating or preventing conditions associated with suppression ofinflammatory responses comprising at least one isoprenyl compound, acarrier and optionally, an additional active ingredient.

In some embodiments, the present invention provides uses of providedcompounds and/or compositions in the treatment of inflammation. Incertain embodiments, the present invention provides uses of providedcompounds and/or compositions in the treatment of diseases that maybenefit from inhibition of infiltration and activation of inflammatorycells (e.g. neutrophils, lymphocytes, monocytes, mast cells), and/orinhibition of expression and activation of cell surface adhesionmolecules (e.g. VCAM-1 and ICAM-1) in endothelial and inflammatorycells. In some embodiments, such includes treating or lessening theseverity of inflammatory diseases or disorders selected frominflammation (acute or chronic), inflammation associated with spinalcord injury to promote nerve regeneration, inhibition of rejection ofgenetically engineered cells by the immune system during in vivo genetherapy, asthma, autoimmune diseases, and chronic obstructive pulmonarydisease (COPD) (e.g., emphysema, chronic bronchitis and small airwaysdisease, etc.), inflammatory responses of the immune system, skindiseases (e.g., reducing acute skin irritation for patients sufferingfrom rosacea, atopic dermatitis, seborrheic dermatitis, psoriasis),irritable bowel syndrome (e.g., Chron's disease and ulcerative colitis,etc.), neurodegenerative disorders (e.g., Parkinson's disease,Alzheimer's disease, Huntington's disease, Dementia pugilistica, Pick'sdisease, Guam parkinsonism dementia complex, Fronto-temporal dementia,Cortico-basal degeneration, Pallido-pontal-nigral degeneration,Progressive supranuclear palsy, Dementia with Lewy bodies (DLB), andmultiple system atrophy (MSA)).

In some embodiments, the present invention provides uses of providedcompounds and/or compositions in medicine, for example, in the treatmentor prevention of diseases or conditions associated with suppression ofinflammatory responses. In certain embodiments, provided herein aremethods for treating or preventing a disease or condition associatedwith suppression of inflammatory responses, methods comprising steps ofadministering an effective amount of a composition comprising at leastone isoprenyl compound, a carrier and optionally an additional activeingredient.

In some embodiments, the present invention provides methods for treatingor preventing diseases in a subject that may benefit from the modulationof levels of inflammatory mediators such as cytokines comprisingprovided compounds and/or compositions. In certain embodiments, thepresent invention provides methods for treating or preventing diseasesin a subject that may benefit from the inhibition of infiltration andaccumulation of helper-T lymphocytes comprising provided compoundsand/or compositions. In certain embodiments, the present inventionprovides methods for treating or preventing diseases in a subject thatmay benefit from the inhibition of ICMT comprising provided compoundsand/or compositions. In certain embodiments, the present inventionprovides methods for treating or preventing diseases in a subject thatmay benefit from inhibition of oxidative burst response from neutrophilscomprising provided compounds and/or compositions.

In some embodiments, provided herein are methods for treating orpreventing skin conditions, said methods comprising the step oftopically applying onto a surface of a subject, including a human, inneed thereof, an effective amount of a composition comprising at leastone isoprenyl compound, a carrier and optionally an additional activeingredient. In certain embodiments, provided herein are methods ofpromoting healthy skin, said methods comprising the step of topicallyapplying onto a surface of a subject, including a human, in needthereof, an effective amount of a composition comprising at least oneisoprenyl compound, a carrier and optionally an additional activeingredient.

In certain embodiments, the present invention provides methods fortreating or preventing inflammation in a subject, methods comprising thestep of administering an effective amount of a composition comprising atleast one isoprenyl compound, a carrier and optionally an additionalactive ingredient.

In some embodiments, provided compounds are compared to AFC. In someembodiments, provided compounds have superior activity to AFC.

While the present specification illustrates certain particular providedcompounds, various other compounds and/or compositions as describedherein would be known to those skilled in the art made aware of thisdisclosure. As described more fully below, in the accompanying figures,examples and descriptions, a related object of this disclosure includesvarious compounds and/or compositions, the choice as to which can bedetermined as desirable for a specific end use application.

DEFINITIONS

“Activating Agent”: As used herein, the term “activating agent” refersto a coupling agent. Exemplary activating agents include, but are notlimited to: benzotriazol-1-yl-oxy-tris-(dimethylamino)-phosphoniumhexafluorphosphate (BOP), N,N′-carbonyldiimidazole (CDI),N,N′-dicyclohhexylcarbodiimide (DCC), N,N′-diisopropylcarbodiimide(DIC), 3-(diethoxyphosphorylloxy)-1,2,3-benzotriazin-4-(3H)-one (DIC),3-(diethoxyphosphorylloxy)-1,2,3-benzotriazin-4-(3H)-one (DEPBT),N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC),2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronoium hexafluorphosphate(HBTU),2-(3,4-dihydro-4-oxo-1,2,3-benzotriazin-3-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HDBTU),2-(mercaptobenzothiazol)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HMTU),2-(endo-5-norbornene-2,3-dicarboxymido)-1,1,3,3-tetramethyluroniumhexafluorophosphate (HNTU), 1-hydroxibenzotriazol monohydrate(HOBt*H2O), 1-hydroxy-1H-1,2,3-Triazole-4-carboxylate (HOCt),N-hydroxy-5-norbornene-2,3-dicarboxylimide (HONB),3-hydroxy-3,4-dihydro-4-oxo-1,2,3-benzotriazine (HOOBt),S-(1-oxido-2-pyridyl)-thio-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HOTT), O-succinimidyl-1,3-dimethylpropyleneuroniumhexafluorophosphate (HPD-OSu),S-(1-oxo-2-pyridyl)-thio-1,3-dimethylpropyleneuroniumhexafluorophosphate (HPTDP),O-(1,2-dihydro-2-oxo-pyridyl]-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HPTU), 2-succinimido-1,1,3,3-tetramethyluroniumhexafluorophosphate (HSTU),4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-morpholinium tetrafluoroborate(MMTM), 1-(mesitylene-2-sulfonyl)-3-nitro-1,2,4-triazole (MSNT),pentafluorphenol-tetramethyluronium hexafluorophosphate (PFTU),tris-n-propan-phosphonic acid anhydride (50% in AcOEt) (PPAA/AcOEt),tris-n-propan-phosphonic acid anhydride (50% in DMF) (PPAA/DMF),2-(1H-benzo triazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU), N,N,N′,N′-tetramethylchloroformamidinium-hexafluorophosphate(TCFH), N,N,N′,N′-tetramethylfluoroformamidinium hexafluorophosphate(TFFH),2-(endo-5-norbornene-2,3-dicarboxymido)-1,1,3,3-tetramethyluroniumtetrafluoroborate (TNTU),S-(1-oxo-2-pyridyl)-thio-N,N,N′,N′-tetramethyluronium tetrafluoroborate(TOTT), O-succinimidyl-1,3-dimethylpropyleneuronium tetrafluoroborate(TPD-OSu), S-(1-oxo-2-pyridyl)-thio-1,3-dimethylpropyleneuroniumtetrafluoroborate (TPTDP),O-(1,2-dihydro-2-oxo-pyridyl]-N,N,N′,N′-tetramethyluroniumtetrafluoroborate (TPTU), orN,N,N′,N′-tetramethyl-O-(succinimidyl)uronium tetrafluoroborate (TSTU),and combinations thereof.

“N-acetyl-S-farnesyl-L-cysteine compound” or an “AFC compound”: As usedherein, an “N-acetyl-S-farnesyl-L-cysteine compound” (or an “AFCcompound”), as used herein, is a small molecule compound that isstructurally related to N-acetyl-S-farnesyl-L-cysteine (AFC). In someembodiments, an AFC compound as provided herein has a structure setforth in Formulae I and/or I′. In some embodiments, an AFC compound mayalso be referred to as an “Isoprenyl Compound.”

“Acyl”: As used herein, the term “acyl” refers to a radical formed froman organic acid by removal of a hydroxyl group.

“Additional active ingredient”: As used herein, the phrase “additionalactive ingredient” refers to an agent, other than an isoprenyl compoundthat exerts a pharmacological, dermatological or any other beneficialactivity. It is to be understood that “other beneficial activity” may beone that is only perceived as such by the subject using the inventivecompositions. Typically, an additional active ingredient, as that termis used herein, refers to a pharmaceutically active agent that isadministered in combination with an isoprenyl compound of the presentinvention.

“Aliphatic”: The term “aliphatic”, as used herein, includes bothsaturated and unsaturated, straight chain (i.e., unbranched), branched,acyclic, cyclic, or polycyclic aliphatic hydrocarbons, which areoptionally substituted with one or more functional groups. As will beappreciated by one of ordinary skill in the art, “aliphatic” is intendedherein to include, but is not limited to, alkyl, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, cycloalkynyl moieties. Thus, as used herein,the term “alkyl” includes straight, branched and cyclic alkyl groups(see below). An analogous convention applies to other generic terms suchas “alkenyl”, “alkynyl”, and the like. Furthermore, as used herein, theterms “alkyl”, “alkenyl”, “alkynyl”, and the like encompass bothsubstituted and unsubstituted groups. In some embodiments, an aliphaticgroup contains 1-25 aliphatic carbon atoms. In some embodiments, analiphatic group contains from 1 to 25, from 1 to 24, from 1 to 23, from1 to 22, from 1 to 21, from 1 to 20, from 1 to 19, from 1 to 18, from 1to 17, from 1 to 16, from 1 to 15, from 1 to 14, from 1 to 13, from 1 to12, from 1 to 11, from 1 to 10, from 1 to 9, from 1 to 8, from 1 to 7,from 1 to 6, from 1 to 5, from 1 to 4, from 1 to 3, from 1 to 2, from 2to 3, or 3 to 4, 4 to 5, 5 to 6, 6 to 7, 7 to 8, 8 to 9, 9 to 10, 10 to11, 11 to 12, 12 to 13, 13 to 14, 14 to 15, 15 to 16, 16 to 17, 17 to18, 18 to 19, 19 to 20, 20 to 21, 21 to 22, 22 to 23, 23 to 24, or 24 to25 aliphatic carbon atoms. In certain embodiments, as used herein,“lower alkyl” is used to indicate those alkyl groups (cyclic, acyclic,substituted, unsubstituted, branched, or unbranched) having 1-6 carbonatoms. In some embodiments, wherein a portion of a term such as alkyl,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl is used withina different generic term (e.g., dialkylamino, alkoxy, alkylthio,alkylamino), then it is understood that an analogous convention applieswith respect to the number of carbon atoms present.

“Alkenyl”: As used herein, the term “alkenyl” denotes a substituted orunsubstituted, monovalent group derived from a straight- orbranched-chain hydrocarbon moiety containing at least one carbon-carbondouble bond by removal of a single hydrogen atom. In some embodiments,the alkenyl group contains 1-25 aliphatic carbon atoms. In certainembodiments, an alkenyl group employed in the invention contains 10-25carbon atoms. In certain embodiments, an alkenyl group employed in theinvention contains 10-20 carbon atoms. In certain embodiments, analkenyl group employed in the invention contains 10-15 carbon atoms. Incertain embodiments, an alkenyl group employed contains 10 carbon atoms.In certain embodiments, an alkenyl group employed contains 15 carbonatoms. In certain embodiments, an alkenyl group employed contains 20carbon atoms. Alkenyl groups include, for example, decenyl, undecenyl,dodecenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl,heptadecenyl, octadecenyl, nonadecenyl, eicosenyl, heneicosenyl,docosenyl, tricosenyl, tetracosenyl, pentacosenyl, polyunsaturatedalkenes including octadec-9,12-dienyl, octadec-9,12,15-trienyl,eicos-5,8,11,14-tetraenyl, farnesyl, geranyl, and geranylgeranyl, C-20phytyl, and the like.

“Alkenylene”: The term “alkenylene” refers to a bivalent, substituted orunsubstituted, alkenyl group. A substituted alkenylene chain is apolymethylene group containing at least one double bond in which one ormore hydrogen atoms are replaced with a substituent. Suitablesubstituents include those described herein for a substituted aliphaticgroup.

“Alkyl”: As used herein, the term “alkyl” means substituted orunsubstituted, saturated, straight- or branched-chain hydrocarbonradicals derived from an aliphatic moiety by removal of a singlehydrogen atom. In some embodiments, the alkyl group contains 1-25aliphatic carbon atoms. In certain embodiments, an alkyl group employedin the invention contains 10-25 carbon atoms. In certain embodiments, analkyl group employed in the invention contains 10-20 carbon atoms. Incertain embodiments, an alkyl group employed in the invention contains15-20 carbon atoms. In certain embodiments, an alkyl group employedcontains 10 carbon atoms. In certain embodiments, an alkyl groupemployed contains 15 carbon atoms. In certain embodiments, an alkylgroup employed contains 20 carbon atoms. In certain embodiments, analkyl group employed in the invention contains 1-3 carbon atoms. Incertain embodiments, an alkyl group employed contains 1-2 carbon atoms.In certain embodiments, an alkyl group contains 1 carbon atom. Examplesof alkyl radicals include, but are not limited to, methyl, ethyl,n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, sec-pentyl,iso-pentyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, sec-hexyl,n-heptyl, n-octyl, n-decyl, n-undecyl, dodecyl, tridecyl, tetradecyl,pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl,heneicosyl, docosyl, tricosyl, teteracosyl, pentacosyl, and the like.

“Alkylamino”: The term “alkylamino” refers to a substituted orunsubstituted group having the structure —NHR′, wherein R′ is aliphatic,as defined herein. In certain embodiments, the aliphatic group contains1-20 aliphatic carbon atoms. In some embodiments, the aliphatic groupcontains 1-10 aliphatic carbon atoms. In some embodiments, the aliphaticgroup employed in the invention contains 1-8 aliphatic carbon atoms. Instill other embodiments, the aliphatic group contains 1-6 aliphaticcarbon atoms. In yet other embodiments, the aliphatic group contains 1-4aliphatic carbon atoms. Examples of alkylamino groups include, but arenot limited to, methylamino, ethylamino, n-propylamino, iso-propylamino,cyclopropylamino, n-butylamino, tert-butylamino, neopentylamino,n-pentylamino, hexylamino, cyclohexylamino, and the like.

“Alkylene”: The term “alkylene” refers to a bivalent substituted orunsubstituted alkyl group. Unless otherwise specified, the alkylenegroup contains 1-25 aliphatic carbon atoms. An “alkylene chain” is apolymethylene group, i.e., —(CH₂)_(n)—, wherein n is a positive integer,preferably from 1 to 6, from 1 to 5, from 1 to 4, from 1 to 3, from 1 to2, from 2 to 3, or 3 to 4, 4 to 5, 5 to 6. A substituted alkylene chainis a polymethylene group in which one or more methylene hydrogen atomsare replaced with a substituent. Suitable substituents include thosedescribed herein for a substituted aliphatic group.

“Alkynyl”: As used herein, the term “alkynyl” denotes a substituted orunsubstituted monovalent group derived from a straight- orbranched-chain hydrocarbon moiety containing at least one carbon-carbontriple bond by removal of a single hydrogen atom. In certainembodiments, an alkynyl group employed in the invention contains 10-25carbon atoms. In certain embodiments, an alkynyl group employed in theinvention contains 10-20 carbon atoms. In certain embodiments, analkynyl group employed contains 10 carbon atoms. In certain embodiments,an alkynyl group employed contains 15 carbon atoms. In certainembodiments, an alkynyl group employed contains 20 carbon atoms. Incertain embodiments, an alkynyl group employed in the invention contains2-3 carbon atoms. In certain embodiments, an alkynyl group employedcontains 2 carbon atoms. In certain embodiments, an alkynyl groupemployed contains 3 carbon atoms. Representative alkynyl groups include,but are not limited to, ethynyl, 2-propynyl (propargyl), 1-propynyl, andthe like.

“Alkoxy”, or “Alkylthio”: The term “alkoxy”, or “alkylthio” as usedherein refers to a substituted or unsubstituted alkyl group, aspreviously defined, attached to the parent molecule through an oxygenatom or through a sulfur atom. In certain embodiments, the “alk” or“alkyl” portion of an “alkoxy” or “alkylthio” group contains 1-10aliphatic carbon atoms. In yet other embodiments, the “alk” or “alkyl”portion of an “alkoxy” or “alkylthio” group employed in the presentinvention contains 1-8 aliphatic carbon atoms. In still otherembodiments, the “alk” or “alkyl” portion of an “alkoxy” or “alkylthio”group contains 1-6 aliphatic carbon atoms. In yet other embodiments, the“alk” or “alkyl” portion of an “alkoxy” or “alkylthio” group contains1-4 aliphatic carbon atoms. Examples of alkoxy, include but are notlimited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, tert-butoxy,neopentoxy, and n-hexoxy. Examples of thioalkyl groups include, but arenot limited to, methylthio, ethylthio, propylthio, isopropylthio,n-butylthio, and the like.

“Animal”: The term animal, as used herein, refers to humans as well asnon-human animals, including, for example, mammals, birds, reptiles,amphibians, and fish. Preferably, the non-human animal is a mammal(e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, aprimate, or a pig). A non-human animal may be a transgenic animal. Insome embodiments, the term animal is used to refer to veterinary animals(e.g., fowl, cows, pigs, horses, etc.).

“Aralkylene” refers to a divalent group of formula —R^(a)—Ar^(a)— whereR^(a) is an “alkylene” as defined herein, and Ar^(a) is an “arylene” asdefined herein (i.e., an alkylene is bonded to an arylene).

“Anti-dandruff agent”: As used herein, the term “anti-dandruff agent” isan agent that reduces, eliminates or prevents a scurf from forming onskin, especially of the scalp, that comes off in small white or grayishscales. Exemplary anti-dandruff ingredients usable in context of thepresent invention include, without limitation, butoconazole, climbazole,coal tar, clotrimazole, dichlorophenyl imidazolodioxalan, imidazoles(e.g., fluconazole, ketoconazole, itraconazole, miconazole,miconazolenitrite, povidone-iodine, sulconazole, tioconazole), salicylicacid, selenium sulfide, shale oil and the like (e.g., sulfonated shaleoil), sulfur, zinc pyrithione, and the like, and any possible stereoisomers thereof such as anthralin, piroctone olamine (Octopirox),selenium sulfide, and ciclopiroxolamine, and combinations thereof.

“Antihistamine agent”: As used herein, the term “antihistamine agent” isan agent that counteracts histamine in the body and that is used fortreating allergic reactions (such as hay fever) and cold symptoms.Non-limiting examples of antihistamines usable in context of the presentinvention include astemizole, brompheniramine, chlorpheniramine,clemastine, dexchlorpheniramine, diphenhydramine, loratadine,piperidines, piperazines, promethazine, terfenadine and tripolidine andcombinations thereof.

“Anti-irritant”: The term “anti-irritant”, as used herein, is an agentthat prevents or reduces soreness, roughness, or inflammation of abodily part (e.g., skin). Presently known anti-irritants can be dividedinto water-soluble anti-irritants and water-insoluble anti-irritants.Representative examples of such compositions are described, for example,in U.S. Pat. No. 5,482,710, which is herein incorporated by reference.Suitable anti-irritants that can be used in the context of the presentinvention include, for example, steroidal and non steroidalanti-inflammatory agents or other materials such as allantoin, aloevera, alpha-bisabolol, caffeine, chamomile, cola nitida extract, greentea extract, glycyrrhizic acid, licorice extract, tea tree oil, or otherxanthines, and combinations thereof.

“Anti-oxidant agent”: As used herein, the term “anti-oxidant agent” isan agent that inhibits oxidation or reactions promoted by oxygen orperoxides. Non-limiting examples of anti-oxidants that are usable in thecontext of the present invention include amines (e.g.,N,N-diethylhydroxylamine, amino-guanidine), arginine pilolate, ascorbicacid (vitamin C) and its salts, ascorbyl esters of fatty acids, ascorbicacid and the like (e.g., magnesium ascorbyl phosphate, sodium ascorbylphosphate, ascorbyl sorbate), bioflavonoids, butylated hydroxy benzoicacids and their salts, curcumin, dihydroxy fumaric acid and its salts,gallic acid and its alkyl esters (e.g., propyl gallate, uric acid andits salts and alkyl esters), glycine pidolate, grape skin/seed extracts,6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (commerciallyavailable under the tradename Trolox®), lipoic acid, lysine, melanin,methionine, nordihydroguaiaretic acid, proline, rosemary extracts,silymarin, sorbic acid and its salts, sulfhydryl compounds (e.g.,glutathione), superoxide dismutase, tea extracts, tocopherol acetate,tocopherol (vitamin E), tocopherol sorbate, and other esters oftocopherol and combinations thereof.

“Antipruritic agents”: As used herein, the term “antipruritic agent” asused herein, is an agent that reduces, eliminates or prevents itching.Suitable antipruritic agents include, without limitation, methdilazineand trimeprazine, and combinations thereof.

“Anti-skin atrophy actives”: As used herein, the term “anti-skin atrophyactive” is an agent that is effective in replenishing or rejuvenatingthe epidermal layer by promoting or maintaining the natural process ofdesquamation. Examples of antiwrinkle and antiskin atrophy actives whichcan be used in context of the present invention include alpha-hydroxyacids (e.g. glycolic acid, and lactic acid), lipoic acid,lysophosphatidic acid, phytic acid, retinoic acid, its prodrugs, isomers(e.g., cis and trans) and analogues thereof, salicylic acid and thelike, sclerosing agents or sclerosants, skin peel agents (e.g., phenoland the like), sulfur-containing D and L amino acids and the like andrelated salts, (e.g., N-acetyl derivatives, such as N-acetylL-cysteine), and thiols (e.g. ethane thiol).

“Anesthetic agents”: The term “anesthetic agent” as used herein is anagent that results in a reduction or loss of sensation. Non-limitingexamples of anesthetic drugs that are suitable for use in the context ofthe present invention include pharmaceutically acceptable salts ofbupivacaine, chlorprocaine, cocaine, dibucaine, dyclonine, etidocaine,hexylcaine, ketamine, lidocaine, mepivacaine, phenol, pramoxine,procaine, and tetracaine.

“Aryl” and “Heteroaryl”: In general, the terms “aryl” and “heteroaryl”refer to substituted or unsubstituted aromatic groups or moieties. Insome embodiments, the terms “aryl” and “heteroaryl” may be used in thecontext of a different moiety name (e.g., “arylalkyl”, “aralkylene”,“aryloxy”, “heteroaryloxy” or “heteroarylalkyl”). In some embodiments,an “aryl” and/or “heteroaryl” refer to stable mono- or polycyclic,heterocyclic, polycyclic, and polyheterocyclic unsaturated moietieswherein at least one ring in the system is aromatic. In someembodiments, an “aryl” and/or “heteroaryl” ring system contains three toseven ring members. In some embodiments, an “aryl” and/or “heteroaryl”contain 3-14 carbon atoms. In certain embodiments of the presentinvention, “aryl” refers to a mono- or bicyclic carbocyclic ring systemhaving one or two aromatic rings including, but not limited to, phenyl,naphthyl, tetrahydronaphthyl, indanyl, indenyl, and the like. In certainembodiments of the present invention, the term “heteroaryl”, as usedherein, refers to a cyclic aromatic radical having from five to ten ringatoms of which one ring atom is selected from S, O, and N; zero, one, ortwo ring atoms are additional heteroatoms independently selected from S,O, and N; and the remaining ring atoms are carbon, the radical beingjoined to the rest of the molecule via any of the ring atoms, such as,for example, pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl,imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl,thiophenyl, furanyl, quinolinyl, isoquinolinyl, and the like. It will beappreciated that aryl and heteroaryl groups can be unsubstituted orsubstituted, wherein substitution includes replacement of one, two,three, or more of the hydrogen atoms thereon independently with any oneor more of the moieties (e.g., “substituents”) provided herein.

“Arylene” and “Heteroarylene”: The term “arylene” refers to anunsubstituted or substituted divalent group that is carbocyclic andaromatic. In some embodiments, rings in an arylene group are fused toone another. In some embodiments rings in an arylene group are notfused, but are nonetheless connected. In some embodiments, an arylenegroup includes some fused rings and some connected rings. In someembodiments, an arylene group includes aromatic rings. In someembodiments, an arylene group includes non-aromatic rings. In someembodiments, an arylene group includes some aromatic rings and somenon-aromatic rings. In some embodiments, the arylene group has up to 5rings, up to 4 rings, up to 3 rings, up to 2 rings, or one aromaticring. For example, the arylene group can be phenylene. Exemplary arylenegroups include any of the “aryl” moieties listed herein with theunderstanding that divalency is required to arrive at a corresponding“arylene” group from an “aryl” group. Exemplary substituents of“arylene” groups include replacement of one, two, three, or more of thehydrogen atoms thereon independently with any one or more of themoieties applicable for “aryl” and “heteroaryl,” as defined herein. Itwill be appreciated by one skilled in the art that a carbon ring atom ofan “arylene” can be replaced by one, two or three heteroatomsindependently selected from S, O, and N while the remaining ring atomsare carbon, the divalent group being joined to the rest of the moleculevia any two ring atoms, to form a “heteroarylene”. Exemplary“heteroarylene” groups include any of the “heteroaryl” moieties listedherein with the understanding that divalency is required to arrive at acorresponding “heteroarylene” group from a “heteroaryl” group.

“Associated with”: When two entities are “associated with” one anotheras described herein, they are linked by a direct or indirect covalent ornon-covalent interaction. Preferably, the association is covalent.Desirable non-covalent interactions include hydrogen bonding, van derWaals interactions, hydrophobic interactions, magnetic interactions,electrostatic interactions, etc.

“Astringent”: As used herein, the term “astringent” is an agent thatdraws together or constricts body tissues and is effective in stoppingthe flow of blood or other secretions. In some embodiments, anastringent coagulate blood, and therefore can be used to arresthemorrhage. In some embodiments, an astringent promotes healing,toughens skin and/or to decreases sweating. In some embodimentsastringents are protein precipitants. Typically, astringents have lowcell penetrability such that their action is limited to the cell surfaceand/or interstitial spaces. In some embodiments, astringent action isaccompanied by contraction and wrinkling of tissues to which astringentsare applied. In some embodiments, application of astringents isaccompanied by blanching of recipient tissue. In some embodiments,astringents include one or more agents such as aluminum, bismuth, iron,manganese, zinc. Alternatively and/or additionally, such agents can beprovided in any of a variety of forms including, for example,pharmaceutically acceptable salt forms.

“Bivalent, branched or unbranched, saturated or unsaturated, C₂-C₆(e.g., C₂, C₃, C₄, C₅, or C₆) hydrocarbon chain”: As used herein, theterm “bivalent, branched or unbranched, saturated or unsaturated, C₂-C₆(e.g., C₂, C₃, C₄, C₅, or C₆) hydrocarbon chain”, refers to bivalentalkylene, alkenylene, and alkynylene chains that are straight orbranched as defined herein.

“Carrier”: The term “carrier” is used in accordance with itsart-understood meaning, to refer to a material that is included in apharmaceutical composition but does not abrogate the biological activityof pharmaceutically active agent(s) that are also included within thecomposition. Typically, carriers have very low toxicity to the animal towhich such compositions are to be administered. In some embodiments,carriers are inert. In some embodiments, carriers are affirmativelybeneficial (e.g., providing pharmaceutical and/or cosmetic benefits). Insome embodiments, isoprenyl compounds of Formulae I, I′ and/or Ia, actas acceptable carriers. In some embodiments, AFC acts as an acceptablecarrier. In some embodiments, the term “carrier” when used in thepharmaceutical context (e.g., pharmaceutically acceptable carrier) meansthat an agent is present in a composition but does not abrogate thebiological activity of another agent(s) present in a composition. Insome embodiments, the term “carrier” when used in a cosmetic context(e.g., cosmetically acceptable carrier) means that an agent is presentin a composition but does not but does not abrogate the biologicalactivity and/or aesthetic effect of another agent(s) present in acomposition. In some embodiments, a cosmetically acceptable carrier isused to topically administer cosmetics with which isoprenyl compounds ofthe present invention will remain stable and bioavailable. It will beunderstood that “cosmetically acceptable carriers” and “carriers” asdefined herein are similar, if not often identical, in nature. In someembodiments, the term “carrier” when used in a cosmeceutical context(e.g., cosmeceutical carrier) means that an agent is present in acomposition but does not abrogate the biological activity and aestheticeffect of another agent(s) present in a composition.

“Caustic agents”: As used herein, the term “caustic agent” is an agentthat is capable of destroying or eating away epithelial tissue bychemical action. Caustic agents can be used to remove dead skin cells.For example, beta-hydroxy acids, naturally derived acids with a strongkerolytic effect, are useful for problem skin or peeling.

“Chelating Agent”: The term “chelating agent” as used herein, is anagent that binds to a metal ion such as calcium (Ca²⁺), magnesium (Mg²⁺)and copper (Cu²⁺), forming a metal complex known as a chelate. In someembodiments, a chelating agent is a ligand. In some embodiments, achelating agent is an atom. In some embodiments, a chelating agent is anion. In some embodiments, a pharmaceutical composition may contain achelating agent (e.g., a mild agent, such as, ethylenediaminetetraaceticacid (“EDTA”), EDTA derivatives, or combinations thereof). In someembodiments, a chelating agent enhances a preservative or preservativesystem of the composition.

“Colorants”: As used herein, the term “colorant” refers to pigmentsand/or dyes or a combination thereof, that are used to change hair coloras cosmetic benefit requires. In some embodiments, pigments included in“colorants” include, but are not limited to, iron oxides, and titaniumoxides. In some embodiments, dyes included in “colorants” include D&Capproved colorants, FD&C approved colorants, and those approved for usein Europe and Japan. See Marmion, D. M., Handbook of US Colorants forFood, Drugs, Cosmetics, and Medical Devices, 3rd ed, 1991 hereinincorporated by reference.

“Compatible”: The term “compatible” as used herein means that thecomponents of such a composition are capable of being combined with eachother in a manner such that there is no interaction that wouldsubstantially reduce the efficacy of the composition under ordinary useconditions.

“Demulcent”: As used herein, the term “demulcent” is an agent used toprimarily alleviate irritation, particularly mucous membranes or abradedtissues. Exemplary demulcents include acacia, agar, alginates,mucilages, benzoin, carbomer, gelatin, glycerin, gums, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose,hydrogels, dextrins, starches, certain sugars, and polymeric polyhydricglycols, propylene glycol, sodium alginate, tragacanth, and combinationsthereof.

“Deodorant agent”: As used herein, the term “deodorant agent” refers toa substance for inhibiting or masking perspiration or other bodilyodors. Representative examples of deodorant agents that are usable inthe context of the present invention include, without limitation,quaternary ammonium compounds such as benzethonium chloride, cetylpyridinium chloride, cetyl-trimethylammonium bromide, diisobutyl phenoxyethoxy ethyl dimethyl benzyl ammonium chloride, lauroyl sarcosine,sodium aluminum chlorohydroxy lactate, sodium N-lauryl sarcosine, sodiumN-palmlthyl sarcosine, N-myristoyl glycine, potassium N-laurylsarcosine, stearyl, trimethyl ammonium chloride, tricetylmethyl ammoniumchloride, 2,4,4′-trichloro-2′-hydroxy diphenyl ether, diaminoalkylamides such as L-lysine hexadecyl amide, heavy metal salts of citrate,salicylate, and piroctose, especially zinc salts, and acids thereof,heavy metal salts of pyrithione, especially zinc pyrithione and zincphenolsulfate. Other deodorant agents include, without limitation, odorabsorbing materials such as carbonate and bicarbonate salts (e.g. as thealkali metal carbonates and bicarbonates, ammonium andtetraalkylammonium carbonates and bicarbonates, especially the sodiumand potassium salts) or combination thereof.

“Dialkylamino”: The term “dialkylamino” refers to a group having thestructure —NRR′, wherein R and R′ are each an aliphatic group, asdefined herein. R and R′ may be the same or different in a dialkylaminomoiety. In certain embodiments, the aliphatic group contains 1-20aliphatic carbon atoms. In some embodiments, the aliphatic groupcontains 1-10 aliphatic carbon atoms. In yet other embodiments, thealiphatic groups employed in the invention contain 1-8 aliphatic carbonatoms. In still other embodiments, the aliphatic group contains 1-6aliphatic carbon atoms. In yet other embodiments, the aliphatic groupcontains 1-4 aliphatic carbon atoms. Examples of dialkylamino groupsinclude, but are not limited to, dimethylamino, methyl ethylamino,diethylamino, methylpropylamino, di(n-propyl)amino, di(iso-propyl)amino,di(cyclopropyl)amino, di(n-butyl)amino, di(tert-butyl)amino,di(neopentyl)amino, di(n-pentyl)amino, di(hexyl)amino,di(cyclohexyl)amino, and the like. In certain embodiments, R and R′ arelinked to form a cyclic structure. The resulting cyclic structure may bearomatic or non-aromatic. Examples of cyclic diaminoalkyl groupsinclude, but are not limited to, aziridinyl, pyrrolidinyl, piperidinyl,morpholinyl, pyrrolyl, imidazolyl, 1,3,4-trianolyl, and tetrazolyl.

“Effective amount”: In general, the “effective amount” of an activeagent (e.g., a therapeutic agent, composition, and/or formulation)refers to an amount sufficient to elicit the desired biologicalresponse. In some embodiments, a therapeutically effective amount of asubstance is an amount that is sufficient, when administered to asubject suffering from or susceptible to a disease, disorder, and/orcondition, to treat, diagnose, prevent, and/or delay the onset of one ormore symptoms of the disease, disorder, and/or condition. As will beappreciated by those of ordinary skill in this art, and effective amountof a substance may vary depending on such factors as the desiredbiological endpoint, the substance to be delivered, the pharmacokineticsof the compound, the target cell or tissue, the disease being treated,the mode of administration, and the patient, etc. For example, theeffective amount of a composition and/or formulation to treat a disease,disorder, and/or condition is the amount that alleviates, ameliorates,relieves, inhibits, prevents, delays onset of, reduces severity ofand/or reduces incidence of one or more symptoms or features of thedisease, disorder, and/or condition. Those of ordinary skill in the artwill appreciate that, commonly, a therapeutically effective amount willbe administered over a series of individual doses. In some embodiments,the term “effective amount” when used in a pharmaceutical context (e.g.,pharmaceutically effective amount) means that an agent is present in anamount sufficient to achieve a desired therapeutic effect. In someembodiments, the term “effective amount” when used in a cosmetic context(e.g., cosmetically effective amount) means that an agent is present inan amount sufficient to achieve an aesthetic effect. In someembodiments, the term “effective amount” when used in a cosmeceuticalcontext (e.g., cosmeceutically effective amount) means that an agent ispresent in an amount sufficient to achieve a therapeutic and/oraesthetic effect.

“Emollients”: As used herein, the term “emollients” refers to an agentthat increases tissue moisture content, thereby rendering skin softerand more pliable. Increased moisture content in the skin can be achievedby preventing water loss with an occlusive water-immiscible barrier, byincreasing the water-holding capacity in the skin with humectants, or byaltering the desquamation of the outermost skin layer, the stratumcorneum. In some embodiments, “emollients” are typically bland, fatty oroleaginous materials which can be applied locally, particularly to theskin. Useful emollients include cetyl alcohol, glycerin, hydrophilicpetrolatum, isopropyl myristate, lanolin, mineral oil, myristyl alcohol,oleyl alcohol, paraffin, petrolatum, spermaceti, vegetable oils, waxes,white ointment, white petroleum, yellow ointment or combinationsthereof.

“Emulsifier”: The term “emulsifier” as used herein promotes formationand stabilization of an emulsion. Suitable emulsifiers may be finelydivided solids, natural materials, or synthetic materials. Naturalemulsifying agents may be derived from either animal or vegetablesources. Those from animal sources include casein, cholesterol, eggyolk, gelatin, or wool fat or combinations thereof. Those from vegetablesources include acacia, chondrus, pectin or tragacanth or combinationsthereof. Vegetable sources specifically from cellulose derivativesinclude carboxymethyl cellulose and methyl cellulose to increase theviscosity. Finely divided emulsifiers include aluminum hydroxide,bentonite, magnesium hydroxide, or magnesium trisylicate. Syntheticagents include anionic, cationic or nonionic agents, and includebenzalkonium chloride, polyethylene glycol 400 monostearate, sodiumlauryl sulfate, or combinations thereof.

“Enantiomerically enriched” and “Enantioenriched”: As used herein, theterms “enantiomerically enriched” and “enantioenriched” denote that oneenantiomer is enriched with respect to other enantiomers of the samecompound in a composition. For example, when a compound is substantiallyin the R-form or the S-form with respect to a particular chiral center,the compound may be considered to have an enantiomeric excess (ee) forthat form. In some embodiments, a composition is considered “enriched”when one enantiomer is present in at least 75% ee in the composition. Incertain embodiments, the terms denote that one enantiomer is present inat least 80% ee, 85% ee, 90% ee, 95% ee, 97.5% ee, or more. In someembodiments, a composition is considered “enantiomerically pure” or“enantiopure” when one enantiomer is present with an ee of at leastabout 90%, with respect to other enantiomers. In some embodiments, acomposition is considered “enantiomerically pure” or “enantiopure” whenone enantiomer is present with an ee of at least about 95%, with respectto other enantiomer(s) present in the composition. In some embodiments,a composition is considered “enantiomerically pure” or “enantiopure”when one enantiomer is present with an ee of at least about 97.5%, withrespect to other enantiomer(s) present in the composition. In someembodiments, a composition is considered “enantiomerically pure” or“enantiopure” when one enantiomer is present with an ee of at leastabout 99%, with respect to other enantiomer(s) present in thecomposition.

“Fragrance”: As used herein, the term “fragrance” refers to an agenthaving a pleasant aroma. Suitable fragrances include, but are notlimited to, camphor synthetic, chamomile, clove oil, eucalyptus oil,lavender, peppermint oil, and the like.

“G-protein mediated condition”: The term “G-protein mediated condition”,as used herein means any disease or other deleterious condition forwhich the appearance, incidence, and/or severity of one or more symptomscorrelates with changes in a G-protein signaling cascade. In someembodiments, one or more symptoms of the disease or condition is causedby a defect or alteration in G-protein signaling.

“Hair Conditioning Agents”: As used herein, the term “hair conditioningagent” refers to an agent that is suitable for use in conditioning hair(e.g., so as to further improve the condition of the hair). In someembodiments, representative hair conditioning agents include, forexample, one or more alkoxylated alcohols, alkoxylated amides,alkoxylated carboxylic acids, cationic surfactants, collagens,dimethicone polyols, esters (e.g., glyceryl esters), halogenatedquaternary ammonium compounds, keratins, modified silicones, proteins,polymeric ethers, quaternary ammonium compounds, or sorbitanderivatives, or combinations thereof.

“Halo” and “Halogen”: The terms “halo” and “halogen” as used hereinrefer to an atom selected from fluorine, chlorine, bromine, and iodine.

“Heteroaliphatic”: The term “heteroaliphatic”, as used herein, refers toaliphatic moieties that contain one or more oxygen, sulfur, nitrogen,phosphorus, or silicon atoms, e.g., in place of carbon atoms.Heteroaliphatic moieties may be branched, unbranched, cyclic or acyclicand include saturated and unsaturated heterocycles such as morpholino,pyrrolidinyl, etc. In certain embodiments, heteroaliphatic moieties aresubstituted by independent replacement of one or more of the hydrogenatoms thereon with one or more moieties (e.g., “substituents”) describedherein.

“Heteroatom”: As used herein, the term “heteroatom” means one or more ofoxygen, sulfur, nitrogen, phosphorus, or silicon (including, anyoxidized form of nitrogen, sulfur, phosphorus, or silicon; thequaternized form of any basic nitrogen or; a substitutable nitrogen of aheterocyclic ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (asin pyrrolidinyl) or NR_(x) (as in N-substituted pyrrolidinyl)).

“Heterocycle” or “Heterocyclyl”: As used herein, the terms“heterocycle”, “heterocyclyl”, “heterocyclic radical”, and “heterocyclicring” are used interchangeably and refer to a stable 3- to 7-memberedmonocyclic or 7-10-membered bicyclic heterocyclic moiety that is eithersaturated or partially unsaturated, and having, in addition to carbonatoms, one or more, preferably one to four heteroatoms independentlyselected from nitrogen, oxygen, or sulfur. When used in reference to aring atom of a heterocycle, the term “nitrogen” includes a substitutednitrogen. As an example, in a saturated or partially unsaturated ringhaving 1-3 heteroatoms selected from oxygen, sulfur or nitrogen, thenitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as inpyrrolidinyl), or NR_(x) (as in N-substituted pyrrolidinyl).

A heterocyclic ring can be attached to its pendant group at anyheteroatom or carbon atom that results in a stable structure and any ofthe ring atoms can be optionally substituted. Examples of such saturatedor partially unsaturated heterocyclic radicals include, withoutlimitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl,piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl,decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl,diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. Theterms “heterocycle”, “heterocyclyl”, “heterocyclyl ring”, “heterocyclicgroup”, “heterocyclic moiety”, and “heterocyclic radical”, are usedinterchangeably herein, and also include groups in which a heterocyclylring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings,such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, ortetrahydroquinolinyl, where the radical or point of attachment is on theheterocyclyl ring. In certain embodiments, one or more carbon atoms maybe substituted with an oxo group in the heterocyclyl ring. Examples ofsuch groups include, without limitation, an isoindolin-1,3-dione moiety.A heterocyclyl group may be mono- or bicyclic. The term“heterocyclylalkyl” refers to an alkyl group substituted by aheterocyclyl, wherein the alkyl and heterocyclyl portions independentlyare optionally substituted.

“Hormone”: As used herein, the term “hormone” refers to naturalsubstances produced by organs of the body that travel by blood totrigger activity in other locations or their synthetic analogs. Suitablehormones for use in the context of the present invention include, butare not limited to, calciferol (Vitamin D₃) and its products, androgens,estrogens and progesterones.

“Hydrocarbon”: The term “hydrocarbon”, as used herein, refers to anychemical group comprising hydrogen and carbon. In some embodiments, ahydrocarbon consists of hydrogen and carbon. A hydrocarbon may besubstituted or unsubstituted. A hydrocarbon may be unsaturated,saturated, branched, unbranched, cyclic, or polycyclic. Illustrativehydrocarbons include, for example, methyl, ethyl, n-propyl, iso-propyl,cyclopropyl, allyl, vinyl, n-butyl, tert-butyl, ethynyl, cyclohexyl,methoxy, diethylamino, and the like. As would be known to one skilled inthis art, all valencies must be satisfied in making any substitutions.As used herein, a “bivalent hydrocarbon” refers to alkylene, alkenylene,or alkynylene, etc.

“Hypopigmenting agents”: As used herein, the term “hypopigmentingagents” refers to substances capable of depigmenting the skin. Suitablehypopigmenting agents include hydroquinones, mequinol, and variousprotease inhibitors including serine protease inhibitors, active soy andretinoic acid.

“In combination”: As used herein, the phrase “in combination” refers toagents that are simultaneously administered to a subject. It will beappreciated that two or more agents are considered to be administered“in combination” whenever a subject is simultaneously exposed to both(or more) of the agents. Each of the two or more agents may beadministered according to a different schedule; it is not required thatindividual doses of different agents be administered at the same time,or in the same composition. Rather, so long as both (or more) agentsremain in the subject's body, they are considered to be administered “incombination”.

“Independently selected”: The term “independently selected” is usedherein to indicate that the R groups can be identical or different.

“Irritant”: As used herein, the term “irritant” is a material that actslocally on the skin to induce, based on irritant concentration,hyperemia, inflammation, and desiccation. Irritant agents include, butare not limited to, alcohol, aromatic ammonia spirits, benzoin tincture,camphor capsicum, and coal tar extracts. In some embodiments, theirritant is a rubefacient.

“Isoprenyl compound”: As used herein, a “isoprenyl compound” is a smallmolecule compound that is structurally related toN-acetyl-S-farnesyl-L-cysteine (AFC) and has the following structure:

wherein L, R¹, R², R³, and Y are as defined herein. In certainembodiments, an isoprenyl compound has the structure set forth inFormula I:

wherein L, R¹, R², R³, and Y are as defined herein. In certainembodiments, an isoprenyl compound has the structure set forth inFormula Ia:

In certain embodiments, an isoprenyl compound has the structure setforth in Formula I′:

In certain embodiments, an isoprenyl compound is(4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid). In certain embodiments, an isoprenyl compound is((E)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobut-2-enoicacid). In certain embodiments, an isoprenyl compound is(4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2-methylene-4-oxobutanoicacid). In certain embodiments an isoprenyl compound is(5-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-5-oxopentanoicacid). In certain embodiments, an isoprenyl compound is5-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2-(1,3-dioxoisoindolin-2-yl)-5-oxopentanoicacid). In certain embodiments, an isoprenyl compound is((R,14E,18E)-15,19,23-trimethyl-4,8-dioxo-3-oxa-12-thia-7,9-diazatetracosa-14,18,22-triene-10-carboxylicacid). In certain embodiments, an isoprenyl compound is((R)-2-(3-(2-carboxyethyl)ureido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid).

“Modulate”: The term “modulate” refers to change in a parameter (e.g., achange in a binding interaction or an activity, etc.). Modulation canrefer to an increase or a decrease in the parameter (e.g., an increaseor decrease in binding, an increase or decrease in activity, etc.).

“Modulator”: The term “modulator” refers to an agent that alters leveland/or activity of its target in an inflammatory pathway. In someembodiments, a modulator alters interaction between a protein in aninflammatory pathway and one or more other entities. In someembodiments, a modulator alters interaction between a protein in aninflammatory pathway and a substrate. Determination of whether an agentis a modulator can be performed directly or indirectly. Determination ofwhether an agent modulates an interaction can be performed directly,e.g., using an assay that detects the interaction between a protein inan inflammatory pathway and a substrate. Determination of whether anagent modulates an interaction can be performed with a technique thatindirectly detects modulation, e.g., a technique that detects abiological activity that is downstream of, and dependent on, theprotein-substrate interaction. In certain embodiments, inflammatorypathways are G-protein-mediated (e.g., purinergic receptor-mediated). Incertain embodiments, inflammatory pathways are non G-protein-mediated(e.g., PPAR-mediated, Toll-like receptor-mediated, and TNF-alphareceptor-mediated).

“Moisturizing agent”: As used herein a “moisturizing agent” is asubstance that adds or restores moisture to the skin. Representativeexamples of moisturizing or humectant agents that are usable in thepresent invention include, without limitation, acetamidemonoethanolamine urazole, aloe vera in any of its variety of forms(e.g., aloe vera gel), allantoin, guanidine, glycolic acid and glycolatesalts (e.g. ammonium salt and quaternary alkyl ammonium salt),hyaluronic acid, lactamide monoethanolamine, polyethylene glycols,polyhydroxy alcohols (e.g., sorbitol, glycerol, hexanetriol, propyleneglycol, butylene glycol, hexylene glycol and the like), sugars andstarches, sugar and starch derivatives (e.g., alkoxylated glucose), andany combination thereof.

“Non-steroidal anti-inflammatory agents”: As used herein, the term“non-steroidal anti-inflammatory agents” refers to a large group ofagents that are aspirin-like in their action, including acetaminophen,Advil®, Aleve®, ibuprofen, naproxen sodium and Tylenol®. Additionalexamples of non-steroidal anti-inflammatory agents that are usable inthe context of the present invention include, without limitation, aceticacid derivatives (e.g., acematacin, clindanac, diclofenac, felbinac,fenclofenac, fentiazac, furofenac, indomethacin, isoxepac, ketorolac,oxepinac, sulindac, tiopinac, tolmetin, zidometacin and zomepirac),benorylate, diflunisal, disalcid, fenamates (e.g., flufenamic,meclofenamic, mefenamic, niflumic and tolfenamic acids), fendosal,oxicams (e.g., CP-14,304, isoxicam, piroxicam, sudoxicarn, andtenoxicam), propionic acid derivatives (e.g., alminoprofen,benoxaprofen, carprofen, fenbufen, fenoprofen, flurbiprofen, ibuprofen,indopropfen, ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen,pranoprofen, suprofen, tiaprofenic and tioxaprofen), pyrazoles (e.g.,azapropazone, feprazone, oxyphenbutazone, phenylbutazone andtrimethazone), safapryn, solprin, trilisate.

“Partially Unsaturated”: As used herein, the term “partiallyunsaturated” refers to a ring moiety that includes at least one doubleor triple bond. The term “partially unsaturated” is intended toencompass rings having multiple sites of unsaturation, but is notintended to include aryl or heteroaryl moieties, as herein defined.

“Penetration enhancer” and “pharmaceutically acceptable penetrationenhancer”: The term “penetration enhancer” and “pharmaceuticallyacceptable penetration enhancer” as used herein is a non-toxic agentthat improves bioavailability of a topical composition. In someembodiments, a penetration enhancer is known to accelerate the deliveryof a substance through the skin (e.g., disrupting the barrier functionof the skin without compromising its barrier effects on microorganismsand toxins). Typically, a penetration enhancer is selected to benon-toxic to skin of the intended recipient (e.g., human). A penetrationenhancer is also desirably compatible with any pharmaceutically activeagent with which it is administered. Representative penetrationenhancers include, for example, and without limitation, such agents as1-substituted azacycloheptane-2-ones (e.g.,1-n-dodecylcyclazacycloheptan-2-one, available under the trademarkAzone® from Whitby Research Incorporated, Richmond, Va.),dipolar-aprotic solvents (e.g., N,N-dimethylacetamide (“DMA”),decylmethylsulfoxide (“C₁₀ MSO”), dimethyl formamide (“DMF”),dimethylsulfoxide (“DMSO”) and N-methyl-2-pyrrolidone (“NMP”)),phospholipids (e.g., allantoin, fatty acid alcohols, lecithin, alcoholsincluding glycerols such as polyethylene glycol monolaurate (“PGML”),glycerol monolaurate (“GML”), urazole, and the like). Penetrationenhancer also can be a vegetable oil, such as, but not limited to, cornoil, cottonseed oil, safflower oil, and olive oil. Additionalpenetration enhancers generally can be found in Remington: The Scienceand Practice of Pharmacy, 20^(th) ed. (Gennaro, A. R., et al., eds.)Lippincott Williams & Wilkins: Philadelphia (2000), which isincorporated herein by reference.

“pH adjusting agent”: As used herein, the term “pH adjusting agent” asused herein is an agent that imparts suitable pH characteristics tocompositions provided herein, (e.g., a substantially neutral pH), the pHof which depends on the specific utilization of the composition. In someembodiments, as the pH of skin is 5.5, it may be desireable to formulatecompositions for topical skin application (to avoid irritation) having apH value in a range of from about 4.0 to about 7.0, or in a range offrom about 5.0 and 6.0, or about 5.5, or substantially 5.5. Suitable pHadjusting agents include, for example, but are not limited to, one ormore adipic acids, buffers, citric acids, calcium hydroxides, glycines,magnesium aluminometasilicates, or combinations thereof.

“Pharmaceutically acceptable salt”: The term “pharmaceuticallyacceptable salt” refers to those salts which are, within the scope ofsound medical judgment, suitable for use in contact with the tissues ofhumans and lower animals without undue toxicity, irritation, allergicresponse, and the like, and are commensurate with a reasonablebenefit/risk ratio. Pharmaceutically acceptable salts are well known inthe art. For example, Berge et al. describe pharmaceutically acceptablesalts in detail in J. Pharmaceutical Sciences, 66: 1-19, 1977;incorporated herein by reference. Such salts can be prepared in situduring the final isolation and purification of the compounds of theinvention, or separately (e.g., by reacting the free base functionalitywith a suitable organic or inorganic acid). Alternatively oradditionally, salts may form during formulation of a compound. Examplesof pharmaceutically acceptable, nontoxic acid addition salts are saltsof an amino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, oxalic acid, maleic acid,tartaric acid, citric acid, succinic acid, or malonic acid or by usingother methods used in the art such as ion exchange. Otherpharmaceutically acceptable salts include adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hernisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate, and aryl sulfonate.

“Pharmaceutically acceptable ester”: The term “pharmaceuticallyacceptable ester” refers to esters which hydrolyze in vivo and includethose that break down readily in the human body to leave the parentcompound or a salt thereof. Suitable ester groups include, for example,those derived from pharmaceutically acceptable aliphatic carboxylicacids, particularly alkanoic, alkenoic, cycloalkanoic, and alkanedioicacids, in which each alkyl or alkenyl moiety advantageously has not morethan 6 carbon atoms. Examples of particular esters include formates,acetates, propionates, butyrates, acrylates, and ethylsuccinates. Incertain embodiments, the esters are cleaved by enzymes such asesterases.

“Pharmaceutically acceptable prodrugs”: The term “pharmaceuticallyacceptable prodrugs” as used herein refers to those prodrugs of thecompounds of the present invention which are, within the scope of soundmedical judgment, suitable for use in contact with the tissues of humansand lower animals with undue toxicity, irritation, allergic response,and the like, commensurate with a reasonable benefit/risk ratio, andeffective for their intended use, as well as the zwitterionic forms,where possible, of the compounds of the invention. The term “prodrug”refers to compounds that are rapidly transformed in vivo to yield theparent compound of the above formula, for example by hydrolysis inblood. A thorough discussion is provided in T. Higuchi and V. Stella,Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S. SymposiumSeries, and in Edward B. Roche, ed., Bioreversible Carriers in DrugDesign, American Pharmaceutical Association and Pergamon Press, 1987,both of which are incorporated herein by reference.

“Preservative”: As used herein, the term “preservative” has itsart-understood meaning and refers to an agent that protects againstundesirable chemical modifications of one or more components in acomposition (e.g., protection against an undesirable chemicalmodification of an active ingredient). Suitable preservatives for use inthe compositions of the present invention include, but are not limitedto, one or more alkanols, disodium EDTA, EDTA salts, EDTA fatty acidconjugates, isothioazolinone, parabens such as methylparaben andpropylparaben, polypropylene glycols, sorbates, urea derivatives such asdiazolindinyl urea, or combinations thereof.

“Propellant”: As used herein, the term“propellant” refers to an agentthat propels the delivery of a composition in, e.g., a vaporized,aerosol nebulized, or spray form. Propellants often are used inmetered-dose inhalers for the treatment of asthma and other respiratorydisorders and for systemic treatments such as insulin for diabetes.Propellants also are used, for example, in nasal inhalers for treatmentof allergic rhinitis, topical sprays, oral sprays, and other aerosolapplications. An example of such propellants, without limitation, arethe Dymel® pharmaceutical propellants manufactured by DuPont™.(Wilmington, Del.).

“Protecting Group”: One of ordinary skill in the art will appreciatethat the synthetic methods, as described herein, utilize a variety ofprotecting groups. By the term “protecting group”, as used herein, it ismeant that a particular functional moiety, e.g., O, S, or N, istemporarily blocked so that a reaction can be carried out selectively atanother reactive site in a multifunctional compound. In preferredembodiments, a protecting group reacts selectively in good yield to givea protected substrate that is stable to the projected reactions; theprotecting group should be selectively removable in good yield byreadily available, preferably non-toxic reagents that do not attack theother functional groups; the protecting group forms an easily separablederivative (more preferably without the generation of new stereogeniccenters); and the protecting group has a minimum of additionalfunctionality to avoid further sites of reaction. As detailed herein,oxygen, sulfur, nitrogen, and carbon protecting groups may be utilized.Hydroxyl protecting groups include methyl, methoxylmethyl (MOM),methylthiomethyl (MTM), t-butylthiomethyl,(phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM),p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM),guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM),siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl,bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR),tetrahydropyranyl (THP), 3-bromotetrahydropyranyl,tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl(MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranylS,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl,2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl,t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl,benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl,p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxido,diphenylmethyl, p,p′-dinitrobenzhydryl, 5-dibenzosuberyl,triphenylmethyl, α-naphthyldiphenylmethyl,p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl,tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxyphenyl)diphenylmethyl,4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl,4,4′,4″-tris(levulinoyloxyphenyl)methyl,4,4′,4″-tris(benzoyloxyphenyl)methyl,3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl,1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl,9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,1,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl(TMS), triethylsilyl (TES), triisopropylsilyl (TIPS),dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS),dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl(TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl,diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate,benzoylformate, acetate, chloroacetate, dichloroacetate,trichloroacetate, trifluoroacetate, methoxyacetate,triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate,3-phenylpropionate, 4-oxopentanoate (levulinate),4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate,adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate,2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate,9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate(TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec),2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutylcarbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkylp-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzylcarbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzylcarbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate,4-ethoxy-1-napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate,4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl,4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,2,6-dichloro-4-methylphenoxyacetate,2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,isobutyrate, mono succinoate, (E)-2-methyl-2-butenoate,o-(methoxycarbonyl)benzoate, α-naphthoate, nitrate, alkylN,N,N′,N′-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate,borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate,sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate(Ts). For protecting 1,2- or 1,3-diols, the protecting groups includemethylene acetal, ethylidene acetal, 1-t-butylethylidene ketal,1-phenylethylidene ketal, (4-methoxyphenyl)ethylidene acetal,2,2,2-trichloroethylidene acetal, acetonide, cyclopentylidene ketal,cyclohexylidene ketal, cycloheptylidene ketal, benzylidene acetal,p-methoxybenzylidene acetal, 2,4-dimethoxybenzylidene ketal,3,4-dimethoxybenzylidene acetal, 2-nitrobenzylidene acetal,methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethyleneortho ester, 1-methoxyethylidene ortho ester, 1-ethoxyethylidine orthoester, 1,2-dimethoxyethylidene ortho ester, α-methoxybenzylidene orthoester, 1-(N,N-dimethylamino)ethylidene derivative,α-(N,N′-dimethylamino)benzylidene derivative, 2-oxacyclopentylideneortho ester, di-t-butylsilylene group (DTBS),1,3-(1,1,3,3-tetraisopropyldisiloxanylidene) derivative (TIPDS),tetra-t-butoxydisiloxane-1,3-diylidene derivative (TBDS), cycliccarbonates, cyclic boronates, ethyl boronate, and phenyl boronate.Amino-protecting groups include methyl carbamate, ethyl carbamate,9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethylcarbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate,2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methylcarbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate(Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate,1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC),1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC),1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2′- and4′-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethylcarbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinylcarbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate(Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc),8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate,2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate,2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methylcarbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc),2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate(Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc),1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate,p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate,2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate,3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methylcarbamate, phenothiazinyl-(10)-carbonyl derivative,N′-p-toluenesulfonylaminocarbonyl derivative, N′-phenylaminothiocarbonylderivative, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzylcarbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentylcarbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate,2,2-dimethoxycarbonylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate,isobutyl carbamate, isonicotinyl carbamate,p-(p′-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate,1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate,1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethylcarbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate,p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate,4-(trimethylammonium)benzyl carbamate, 2,4,6-trimethylbenzyl carbamate,formamide, acetamide, chloroacetamide, trichloroacetamide,trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide,3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide,p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide,acetoacetamide, (N′-dithiobenzyloxycarbonylamino)acetamide,3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide,2-methyl-2-(o-nitrophenoxy)propanamide,2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethioninederivative, o-nitrobenzamide, o-(benzoyloxymethyl)benzamide,4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts),N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole,N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE),5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine,N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl)amine, quaternary ammoniumsalts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),N-9-phenylfluorenylamine (PhF),N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm),N-2-picolylamino N′-oxide, N-1,1-dimethylthiomethyleneamine,N-benzylideneamine, N-p-methoxybenzylideneamine,N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,N—(N′,N′-dimethylaminomethylene)amine, N,N′-isopropylidenediamine,N-p-nitrobenzylideneamine, N-salicylideneamine,N-5-chlorosalicylideneamine,N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,N-borane derivative, N-diphenylborinic acid derivative,N-[phenyl(pentacarbonylchromium- or tungsten)carbonyl]amine, N-copperchelate, N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide,diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt),diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzylphosphoramidate, diphenyl phosphoramidate, benzenesulfenamide,o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide,pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide,triphenylmethylsulfenamide, 3-nitropyridinesulfenamide (Npys),p-toluenesulfonamide (Ts), benzenesulfonamide,2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mtr),2,4,6-trimethoxybenzenesulfonamide (Mtb),2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide(Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide(Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide,4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide,phenyl urea, ethylurea and cyclopropyl sulfonamide. Exemplary protectinggroups are detailed herein. However, it will be appreciated that thepresent invention is not intended to be limited to these protectinggroups; rather, a variety of additional equivalent protecting groups canbe readily identified using the above criteria and utilized in themethod of the present invention. Additionally, a variety of protectinggroups are described in Protective Groups in Organic Synthesis, ThirdEd. Greene, T. W. and Wuts, P. G., Eds., John Wiley & Sons, New York:1999, the entire contents of which are hereby incorporated by reference.

“Protective”: As used herein, the term “protective” refers to an agentthat isolates exposed surface of skin or other membrane from harmful orannoying stimuli. Exemplary protectives include dusting powders,adsorbents, mechanical protective agents, and plasters. Mechanicalprotectives are generally either collodions or plasters, and include,for example aluminum hydroxide gel, collodium, dimethicone, petrolatumgauze, absorbable gelatin film, absorbable gelatin sponge, zinc gelatin,kaolin, lanolin, anhydrous lanolin, mineral oil, mineral oil emulsion,mineral oil light, olive oil, peanut oil, petrolatum, silicones,hydrocolloids and the like. In some embodiments, a protective includesan adherent, continuous film that may be flexible or semi-rigiddepending on the materials and the formulations as well as the manner inwhich they are applied. In some embodiments, a “protective” may be a“demulscent” as described herein.

“Racemic”: As used herein, a racemic mixture means about 50% of oneenantiomer and about 50% of its corresponding enantiomer relative to allchiral centers in a molecule. Compounds of the present invention mayencompass enantiomerically pure, enantiomerically enriched, and racemicmixtures.

“Rubefacient”: As used herein, the term “rubefacient” is an agent thatinduces hyperemia, wherein hyperemia means an increased amount of bloodin a body part or organ. Rubefaction, which is induced by rubefacients,results from increased circulation to an injured area and is accompaniedby a feeling of comfort, warmth, itching and hyperesthesia.

“Sclerosant”: As used herein, the term “sclerosant” is an agent (e.g.,chemical irritant) that is injected into a vein in sclerotherapy.Exemplary sclerosants include laureth 9 and ethanolamine oleate,morrhuate sodium, sodium tetradecyl sulfate.

“Skin Irritant”: As used herein, the term “skin irritant” refers to acompound that, when applied to skin or a skin equivalents, elicits acellular response characterized by the expression of an “irritantresponsive gene.” Examples of known skin irritants include, but are notlimited to, sodium dodecyl sulfate (“SDS”), calcipotriol, andtrans-retinoic acid. The term “skin irritant” is also intended toencompass unknown or suspected irritants, including but not limited to,those containing in some pharmaceuticals, cosmetics, and consumerproducts.

“Small Molecule”: As used herein, the term “small molecule” refers to anorganic compound either synthesized in the laboratory or found innature. Typically, a small molecule is characterized in that it containsseveral carbon-carbon bonds, and has a molecular weight of less than1500, although this characterization is not intended to be limiting forthe purposes of the present invention. Examples of “small molecules”that occur in nature include, but are not limited to, taxol, dynemicin,and rapamycin. Examples of “small molecules” that are synthesized in thelaboratory include, but are not limited to, the inventive compoundsincorporated herein.

“Solubilizing agent”: As used herein, the term “solubilizing agent” arethose substances that enable solutes to dissolve. Representativeexamples of solubilizing agents that are usable in the context of thepresent invention include, without limitation, complex-formingsolubilizers (e.g., citric acid, ethylenediamine-tetraacetate, sodiummeta-phosphate, succinic acid, urea, cyclodextrin, polyvinylpyrrolidone,diethylammonium-ortho-benzoate, etc.), n-alkyl amine n-oxides,micelle-forming solubilizers (e.g., TWEEN®, including TWEEN 80®),organic solvents (e.g., acetone, phospholipids and cyclodextrins),polyoxamers, polyoxyethylene n-alkyl ethers, and polyoxyethylenesorbitan fatty acid ester.

“Steroidal anti-inflammatory agent”: As used herein, the term “steroidalanti-inflammatory agent”, refers to any one of numerous compoundscontaining a 17-carbon 4-ring system and includes the sterols, varioushormones (as anabolic steroids), and glycosides. Representative examplesof steroidal anti-inflammatory drugs include, without limitation,cortico steroids such as alpha-methyl dexamethasone, amcinafel,amcinafide, beclomethasone dipropionates, beclomethasone dipropionate,betamethasone and the balance of its esters, chloroprednisone,chlorprednisone acetate, clescinolone, clobetasol valerate,clocortelone, cortisone, cortodoxone, desonide, desoxycorticosteroneacetate, desoxymethasone, dexamethasone, dexamethasone-phosphate,dichlorisone, dichlorisone, diflorasone diacetate, diflucortolonevalerate, difluorosone diacetate, diflurosone diacetate, diflurprednate,fluadrenolone, flucetonide, fluclorolone acetonide, flucloronide,flucortine butylesters, fludrocortisone, fludrocortisone,fludrocortisone, flumethasone pivalate, flunisolide, fluocinonide,fluocortolone, fluoromethalone, fluosinolone acetonide, fluperolone,fluprednidene (fluprednylidene) acetate, fluprednisolone, fluradrenoloneacetonide, fluradrenolone, flurandrenolone, halcinonide, hydrocortamate,hydrocortisone acetate, hydrocortisone butyrate, hydrocortisonevalerate, hydrocortisone cyclopentylpropionate, hydrocortisone,hydroxyltriamcinolone, medrysone, meprednisone, methylprednisolone,paramethasone, prednisolone, prednisone, triamcinolone acetonide,triamcinolone, and combinations thereof.

“Substituted”: It will be appreciated that the compounds, as describedherein, may be substituted with any number of substituents or functionalmoieties. In general, the term “substituted” whether preceded by theterm “optionally” or not, and substituents contained in formulas of thisinvention, refer to the replacement of hydrogen radicals in a givenstructure with the radical of a specified substituent. When more thanone position in any given structure may be substituted with more thanone substituent selected from a specified group, the substituent may beeither the same or different at every position. As used herein, the term“substituted” is contemplated to include all permissible substituents oforganic compounds. In a broad aspect, the permissible substituentsinclude acyclic and cyclic, branched and unbranched, carbocyclic andheterocyclic, aromatic and nonaromatic substituents of organiccompounds. Heteroatoms such as nitrogen may have hydrogen substituentsand/or any permissible substituents of organic compounds describedherein which satisfy the valencies of the heteroatoms. Furthermore, thisinvention is not intended to be limited in any manner by the permissiblesubstituents of organic compounds. Combinations of substituents andvariables envisioned by this invention are preferably those that resultin the formation of stable compounds useful in the treatment, forexample, of inflammatory diseases and/or disorders, e.g., in themodulation of a G-protein signaling cascade.

Some examples of substituents of aliphatic and other moieties ofcompounds provided by the present invention include, but are not limitedto aliphatic; heteroaliphatic; aryl (e.g., phenyl); heteroaryl;arylalkyl; heteroarylalkyl; alkoxy; aryloxy; arylthio, heteroalkoxy;heteroaryloxy; alkylthio; arylthio; heteroalkylthio; heteroarylthio; —F;—Cl; —Br;

—I; —OH; —NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH; —CH₂NH₂;—CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x); —NR_(x)(CO)R_(x)wherein each occurrence of R_(x) independently includes, but is notlimited to, aliphatic, heteroaliphatic, aryl, heteroaryl, arylalkyl, orheteroarylalkyl, wherein any of the aliphatic, heteroaliphatic,arylalkyl, or heteroarylalkyl substituents described above and hereinmay be substituted or unsubstituted, branched or unbranched, cyclic oracyclic, and wherein any of the aryl or heteroaryl substituentsdescribed above and herein may be substituted or unsubstituted.Additional examples of generally applicable substituents are illustratedby the specific embodiments described herein.

“Stable”: As used herein, the term “stable” preferably refers to thestate of maintaining integrity of a compound over a period of time(e.g., during manufacture and/or storage).

“Substantially free of”: As used herein, the term “substantially freeof”, when used to describe a material or compound, means that thematerial or compound lacks a significant or detectable amount of adesignated substance. In some embodiments, the designated substance ispresent at a level not more than about 1%, 2%, 3%, 4% or 5% (w/w or v/v)of the material or compound.

“Surfactants”: As used herein, the term “surfactant” is a surface-activesubstance, such as a detergent. Suitable surfactants for use with theinventive compositions include, but are not limited to, sarcosinates,glutamates, sodium alkyl sulfates, ammonium alkyl sulfates, sodiumalkyleth sulfates, ammonium alkyleth sulfates, ammoniumlaureth-n-sulfates, sodium laureth-n-sulfates, isothionates,glycerylether sulfonates, sulfosuccinates and combinations thereof. Moreparticularly, an anionic surfactant is selected from the groupconsisting of sodium lauroyl sarcosinate, monosodium lauroyl glutamate,sodium alkyl sulfates, ammonium alkyl sulfates, sodium alkylethsulfates, ammonium alkyleth sulfates, and combinations thereof.

“Sun screening agent”: As used herein, a “sun screening agent” refers toan agent, when topically applied, absorbs or reflects some of the sun'sultraviolet radiation on skin exposed to sunlight, and therefore helpsprotect against sunburn. In some embodiments, a sun screening agentabsorbed in the skin may lead to an increase in reactive oxygen species.Representative examples of sun screening agents usable in the presentinvention include, without limitation, p-aminobenzoic acid and its saltsand derivatives thereof (p-dimethylaminobenzoic acid; ethyl, glyceryl,and isobutyl esters); anthranilates (i.e., o-amino-benzoates; benzyl,cyclohexenyl, linalyl, menthyl, methyl, phenyl, phenylethyl, andterpinyl esters); benzophenones (i.e., hydroxy- or methoxy-substitutedbenzophenones such as benzoresorcinol, butylmethoxydibenzoylmethane,2,2′-dihydroxy-4,4′-dimethoxybenzophenone, etocrylene,4-isopropyldibenzoylmethane, dioxybenzone,3-4′-methylbenzylidene-boman-2-one, octabenzone, octocrylene,oxybenzene, sulisobenzone, and 2,2′,4,4′-tetrahydroxybenzophenone);(butyl carbotol) (6-propyl piperonyl) ether; cinnamic acid derivatives(alpha.-phenyl cinnamonitrile; butyl cinnamoyl pyruvate; benzyl andmethyl esters); diazoles (2-acetyl-3-bromoindazole, aryl benzothiazoles,methyl naphthoxazole, and phenyl benzoxazole); dibenzylacetone;dihydroxycinnamic acid derivatives (methylaceto-umbelliferone,methylumbelliferone, umbelliferone); di-hydroxynaphthoic acid and itssalts; hydrocarbons (diphenylbutadiene, and stilbene); hydroquinone; o-and p-hydroxybiphenyldisulfona-tes; coumarin derivatives (3-phenyl,7-hydroxy, and 7-methyl); naphtholsulfonates (sodium salts of2-naphthol-3,6-disulfonic and of 2-naphthol-6,8-disulfonic acids);quinine salts (bisulfate, chloride, oleate, sulfate and tannate);quinoline derivatives (8-hydroxyquinoline salts, and 2-phenylquinoline);salicylates (amyl, benzyl, di-propylene glycol, glyceryl, menthyl,octyl, and phenyl esters); tannic acid and its derivatives (e.g.,hexaethylether); trihydroxy-cinnamic acid derivatives (daphnetin,daphnin, esculetin, esculin, methylesculetin; and the glucosides); anduric and violuric acids; and combinations thereof.

“Thickeners”: As used herein, the term “thickener” refers to agents thatmake a composition more dense or viscous in consistency. Suitablethickeners that can be used in the context of the present inventioninclude, for example, non-ionic water-soluble polymers such ashydroxyethylcellulose (commercially available under the TrademarkNatrosol® 250 or 350), cationic water-soluble polymers such as Polyquat37 (commercially available under the Trademark Synthalen® CN), fattyalcohols, fatty acids, anionic polymers, and their alkali salts andmixtures thereof.

“Thio”: As used herein, the term “thio” used alone or as part of alarger moiety as in “alkylthio”, “arylthio”, “heteroalkylthio”, or“heteroarylthio” refers to replacement of an oxygen. For example,“alkylthio” refers to an alkyl group, as previously defined, attached tothe parent molecule through a sulfur atom. Similarly, “arylthio” refersto an aryl group, as previously defined, attached to the parent moleculethrough a sulfur molecule. Similarly, “heteroalkylthio” refers to aheteroalkyl group, as previously defined, attached to the parentmolecule through a sulfur molecule, etc.

“Treat,” “treating” and “treatment”: As used herein, the terms “treat,”“treating” and “treatment,” contemplate an action that occurs while apatient is suffering from or susceptible to a specified disease,disorder or condition, which delays onset of and/or reduces thefrequency or severity of one or more symptoms or features of the diseasedisorder or condition. Thus, “treat”, “treating”, and “treatment” referto any type of treatment that imparts a benefit to a subject afflictedwith a disease, disorder or condition, including improvement in thecondition of the subject (e.g., in one or more symptoms), delay in theprogression of the disease, disorder or condition, prevention or delayof the onset of the disease, disorder or condition, etc.

Unit dosage form: The expression “unit dosage form” as used hereinrefers to a physically discrete unit of a provided formulationappropriate for the subject to be treated. It will be understood,however, that the total daily usage of provided formulation will bedecided by the attending physician within the scope of sound medicaljudgment. The specific effective dose level for any particular subjector organism may depend upon a variety of factors including the disorderbeing treated and the severity of the disorder; activity of specificactive agent employed; specific formulation employed; age, body weight,general health, sex and diet of the subject; time of administration, andrate of excretion of the specific active agent employed; duration of thetreatment; drugs and/or additional therapies used in combination orcoincidental with specific compound(s) employed, and like factors wellknown in the medical arts. In some embodiments, a unit dosage formcontains an amount of a therapeutically active agent appropriate for usein a therapeutic regimen (i.e., in a regimen that delivers atherapeutically effective amount of an agent). In some embodiments, sucha unit dosage form may be considered to contain a “therapeuticallyeffective amount” of an agent even if a single dose would not beexpected to be effective.

“Unsaturated”: As used herein, the term “unsaturated” means that amoiety has one or more units of unsaturation.

“Vitamin”: As used herein, the term “vitamin” refers to any of variousorganic substances essential in minute quantities to the nutrition ofmost animals act especially as coenzymes and precursors of coenzymes inthe regulation of metabolic processes. Non-limiting examples of vitaminsusable in context of the present invention include vitamin A and itsanalogs and derivatives: retinol, retinal, retinyl palmitate, retinoicacid, tretinoin, iso-tretinoin (known collectively as retinoids),vitamin E (tocopherol and its derivatives), vitamin C (L-ascorbic acidand its esters and other derivatives), vitamin B₃ (niacinamide and itsderivatives), alpha hydroxy acids (such as glycolic acid, lactic acid,tartaric acid, malic acid, citric acid, etc.) and beta hydroxy acids(such as salicylic acid and the like).

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 is a table depicting % inhibition determined from an edema assay,an erythema assay, and myeloperoxidase (“MPO”) assay for compound A,compound B, compound C, compound D, compound E, compound F, compound Gand AFC.

FIG. 2 is a table depicting ED₅₀ results (μg/ear) obtained for AFC,Compound A and Compound B using an edema assay, an erythema assay, andmyeloperoxidase (“MPO”) assay, as described infra.

FIG. 3 is a table summarizing activity ranges determined from an MPOactivity assay for exemplary compounds in Table 1.

FIG. 4A and FIG. 4B are bar graphs depicting ED₅₀ results (μg/ear)obtained for Compound A, demonstrating that administering Compound A at0.25%, 0.50% and 1.0% dosage levels results in an inhibition in TNF-α(FIG. 4A panel) and IL-1β (FIG. 4B panel) levels, as determined using aTPA mouse ear inflammation model.

FIG. 5A and FIG. 5B depict bar graphs depicting IL-8 levels (pg/mL)obtained for Compound A in the presence (FIG. 5A panel) and absence(FIG. 5B panel) demonstrating a dose dependent inhibition of LPS-TLR4induced IL-8 release with, as determined using human microvascularendothelial cell line-1 (HMEC-1) cultures.

FIG. 6A and FIG. 6B are bar graphs depicting IL-8 levels (pg/mL)obtained for Compound A in the presence (FIG. 6A panel) and absence(FIG. 6B panel) of ATP-γS, demonstrating a dose dependent inhibition ofATP-γS-purinergic receptor-induced IL-8 release, as determined usinghuman microvascular endothelial cell line-1 (HMEC-1) cultures.

FIG. 7A and FIG. 7B are bar graphs depicting MCP-1 levels (pg/mL)obtained for Compound A in the presence (FIG. 7A panel) and absence(FIG. 7B panel) of ATP-γS, demonstrating a dose dependent inhibition ofATP-γS-purinergic receptor-induced IL-8 release, as determined usinghuman microvascular endothelial cell line-1 (HMEC-1) cultures.

FIG. 8A and FIG. 8B are bar graphs depicting IL-8 levels (pg/mL)obtained for Compound A, demonstrating a dose dependent inhibition ofTPA-induced IL-8 release, as determined using Normal Human EpidermalKeratinocyte (NHEK) cell cultures.

FIG. 9 is a graph depicting IL-8 levels (pg/mL) obtained for AFC,Compound A and Compound B, demonstrating a dose dependent inhibition ofTNF-alpha induced IL-8 release, as determined using Human Umbilical VeinEndothelial cell (HUVEC) cultures.

FIG. 10 is a treatment protocol for administering Compound B in aK5.Stat3c psoriasis mouse model.

FIG. 11 is a bar graph depicting number of CD3+ cells/mm skin obtainedwith Compound B, demonstrating a dose dependent inhibition of number ofCD3+ (Helper-T-lymphocytes), as determined using a transgenic mousemodel (K5.Stat3c) for psoriasis.

FIG. 12 is a bar graph depicting % inhibition of the G-proteinmethylating enzyme ICMT obtained for compound N-64, Compound N-19,Compound A, Compound N-30 and Compound N-77, as demonstrated by the %reduction of methylated acetyl-farnesyl-cysteine, an ICMT substrate.

FIG. 13 is a bar graph depicting % inhibition of oxidative burst fromneutrophils obtained for AFC, Compound C, Compound N-25, AFC-methylester (AFC-ME) and AFC-AcetoxylMethane (AFC-AM), as demonstrated by %reduction of superoxide formation.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS 1. Description of ExemplaryCompounds

Compounds provided by the present invention include those describedgenerally above, and are further illustrated by all classes, subclassesand species of each of these compounds disclosed herein.

According to one aspect, the present invention provides compounds ofFormula:

or a pharmaceutically acceptable salt thereof, wherein:

L is a bivalent, branched or unbranched, saturated or unsaturated, C₂-C₆hydrocarbon chain wherein one or more methylene units of L isindependently replaced by —O—, —S—, —NH—, —C(O)—, —C(═CH₂)—, or C₃-C₆cycloalkylene, wherein L is optionally substituted by one or more groupsselected from halogen, phenyl, an 8-10 membered bicyclic aryl ring, a5-6 membered heteroaryl ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, an 8-10 membered bicyclicheteroaryl ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, or a 5- to 7-membered monocyclic or 7-10membered bicyclic heterocyclyl ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur;

R¹ is hydrogen, —OH or —OR, wherein each R is independently hydrogen, anoptionally substituted group selected from C₁₋₆ aliphatic or C₁₋₆heteroaliphatic, —NHR, —NH(OR), —ONH₂, or —NR₂;

R² is —C(O)X, wherein X is independently R, —OR, a hydrogen, aryloxy,amino, alkylamino, dialkylamino, heteroaryloxy, hydrazine, a 6-10membered aryl ring, a 5-6 membered heteroaryl ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein each R is independently hydrogen or an optionally substitutedgroup selected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic;

R³ is a substituted or unsubstituted, branched or unbranched, saturatedor unsaturated, C₁₀-C₂₅ aliphatic; and

Y is —O—, —N—, —S—, —Se—, —S(O)—, —S(═N)—, —SO₂—, —Se(O)—, or —Se(O)₂—.

In some embodiments, compounds of the above-described Formula areprovided with the proviso that L and R¹ cannot together be C₁-C₃unsubstituted non-halogenated alkyl.

In some embodiments, the present invention provides a compound ofFormula I′:

or a pharmaceutically acceptable salt thereof, wherein:

L is a bivalent, branched or unbranched, saturated or unsaturated, C₂-C₆hydrocarbon chain wherein one or more methylene units of L isindependently replaced by —O—, —S—, —NH—, —C(O)—, —CF₂—, —C(═CH₂)—,—CH═CH—, or an optionally substituted arylene, heteroarylene, C₃-C₆cycloalkylene, C₃-C₆ heterocycloalkylene, or an 8-10-membered bicyclicheterocyclic moiety,

and wherein L is optionally substituted by one or more groups selectedfrom halogen, C₁-C₆ alkyl, phenyl, biphenyl, -benzyl, —CH₂-phenol,—CH(phenyl)₂, —OH, —NH₂, —NHC(O)CH₃, —NHC(O)NHCH₂CH₃, —C(O)NH₂,—C(O)NHCH₂CH₃, —CH₂C(O)OCH₂phenyl, —(CH₂)₂SCH₃, —(CH₂)₂C(O)NH₂,—(CH₂)₂C(O)OH, an 8-10 membered bicyclic aryl ring, a 5-6 memberedheteroaryl ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, an 8-10 membered bicyclic heteroaryl ringhaving 1-4 heteroatoms independently selected from nitrogen, oxygen, orsulfur, a 5- to 7-membered monocyclic having 1-2 heteroatomsindependently selected from nitrogen, oxygen, or sulfur or a 7-10membered bicyclic heterocyclyl ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur;

M is —C(O)—, —C(S), or —SO₂—;

R¹ is hydrogen, F, CF₃, C₁-C₄ alkyl, —OH, —C(O)CH₃, —NH(OR), —NR₂,—NHNR₂, —SO₂R, —NH-phenyl, —SO₂-phenyl, -phenyl-NO₂, or —OR, whereineach R is independently hydrogen, oxygen, or an optionally substitutedgroup selected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic;

R² is —C(O)X, wherein X is independently R, —C(O)NHNH₂, —OR, a hydrogen,aryloxy, amino, alkylamino, dialkylamino, heteroaryloxy, hydrazine, a6-10 membered aryl ring, a 5-6 membered heteroaryl ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein each R is independently hydrogen or an optionally substitutedgroup selected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic; and

R³ is a substituted or unsubstituted, branched or unbranched, saturatedor unsaturated, C₁₀-C₂₅ aliphatic; and

Y is —O—, —N—, —S—, —Se—, —S(O)—, —S(═N)—, —SO₂—, —Se(O)—, or —Se(O)₂—.

In some embodiments, compounds of Formula I′ are provided with theproviso that L and R¹ cannot together be C₁-C₃ unsubstitutednon-halogenated alkyl.

According to one aspect, the present invention provides compounds ofFormula I,

or a pharmaceutically acceptable salt thereof, wherein:

L is a bivalent, branched or unbranched, saturated or unsaturated, C₂-C₆hydrocarbon chain wherein one or more methylene units of L isindependently replaced by —O—, —S—, —NH—, —C(O)—, —C(═CH₂)—, or C₃-C₆cycloalkylene, wherein L is optionally substituted by one or more groupsselected from halogen, phenyl, an 8-10 membered bicyclic aryl ring, a5-6 membered heteroaryl ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, an 8-10 membered bicyclicheteroaryl ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, or a 5- to 7-membered monocyclic or 7-10membered bicyclic heterocyclyl ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur;

R¹ is hydrogen, —OH or —OR, wherein each R is independently hydrogen oran optionally substituted group selected from C₁₋₆ aliphatic or C₁₋₆heteroaliphatic;

R² is —C(O)X, wherein X is independently R, —OR, a hydrogen, aryloxy,amino, alkylamino, dialkylamino, heteroaryloxy, hydrazine, a 6-10membered aryl ring, a 5-6 membered heteroaryl ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein each R is independently hydrogen or an optionally substitutedgroup selected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic; and

R³ is a substituted or unsubstituted, branched or unbranched, saturatedor unsaturated, C₁₀-C₂₅ aliphatic.

According to another aspect, the present invention provides compounds ofFormula I,

or a pharmaceutically acceptable salt thereof, wherein:

L is a bivalent, branched or unbranched, saturated or unsaturated, C₂-C₆hydrocarbon chain wherein one or more methylene units of L isindependently replaced by —O—, —S—, —NH—, —C(O)—, —C(═CH₂)—, or C₃-C₆cycloalkylene, wherein L is optionally substituted by one or more groupsselected from halogen, phenyl, an 8-10 membered bicyclic aryl ring, a5-6 membered heteroaryl ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, an 8-10 membered bicyclicheteroaryl ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, a 5- to 7-membered monocyclic having 1-2heteroatoms independently selected from nitrogen, oxygen, or sulfur or a7-10 membered bicyclic heterocyclyl ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, or sulfur;

R¹ is hydrogen, —OH or —OR, wherein each R is independently hydrogen oran optionally substituted group selected from C₁₋₆ aliphatic or C₁₋₆heteroaliphatic;

R² is —C(O)X, wherein X is independently R, —OR, a hydrogen, aryloxy,amino, alkylamino, dialkylamino, heteroaryloxy, hydrazine, a 6-10membered aryl ring, a 5-6 membered heteroaryl ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein each R is independently hydrogen or an optionally substitutedC₁₋₆ aliphatic; and

R³ is a substituted or unsubstituted, branched or unbranched, saturatedor unsaturated, C₁₀-C₂₅ aliphatic.

In certain embodiments, the present invention provides a compound ofFormula Ia,

or a pharmaceutically acceptable salt thereof, wherein:

L is a bivalent, branched or unbranched, saturated or unsaturated, C₂-C₆hydrocarbon chain wherein one or more methylene units of L isindependently replaced by —O—, —S—, —NH—, —C(O)—, —C(═CH₂)—, or C₃-C₆cycloalkylene, wherein L is optionally substituted by one or more groupsselected from halogen, phenyl, an 8-10 membered bicyclic aryl ring, a5-6 membered heteroaryl ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, an 8-10 membered bicyclicheteroaryl ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, a 5- to 7-membered monocyclic having 1-2heteroatoms independently selected from nitrogen, oxygen, or sulfur or a7-10 membered bicyclic heterocyclyl ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, or sulfur;

R¹ is hydrogen, —OH or —OR, wherein each R is independently hydrogen oran optionally substituted group selected from C₁₋₆ aliphatic or C₁₋₆heteroaliphatic; and

R² is —C(O)X, wherein X is independently R, —OR, a hydrogen, aryloxy,amino, alkylamino, dialkylamino, heteroaryloxy, hydrazine, a 6-10membered aryl ring, a 5-6 membered heteroaryl ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein each R is independently hydrogen or an optionally substitutedgroup selected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic.

As defined generally above, the L group of Formulae I, and/or I′ is abivalent, branched or unbranched, saturated or unsaturated, C₂-C₆hydrocarbon chain wherein one or more methylene units of L isindependently replaced by —O—, —S—, —NH—, —C(O)—, —CF₂—, —C(═CH₂)—,—CH═CH—, or an optionally substituted arylene, heteroarylene, C₃-C₆cycloalkylene, C₃-C₆ heterocycloalkylene, or an 8-10-membered bicyclicheterocyclic moiety,

and wherein L is optionally substituted by one or more groups selectedfrom halogen, C₁-C₆ alkyl, phenyl, biphenyl, -benzyl, —CH₂-phenol,—CH(phenyl)₂, —OH, —NH₂, —NHC(O)CH₃, —NHC(O)NHCH₂CH₃, —C(O)NH₂,—C(O)NHCH₂CH₃, —CH₂C(O)OCH₂phenyl, —(CH₂)₂SCH₃, —(CH₂)₂C(O)NH₂,—(CH₂)₂C(O)OH, an 8-10 membered bicyclic aryl ring, a 5-6 memberedheteroaryl ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, an 8-10 membered bicyclic heteroaryl ringhaving 1-4 heteroatoms independently selected from nitrogen, oxygen, orsulfur, a 5- to 7-membered monocyclic having 1-2 heteroatomsindependently selected from nitrogen, oxygen, or sulfur or a 7-10membered bicyclic heterocyclyl ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur.

Particular embodiments of different moieties/groups included incompounds of the present invention are discussed in more detail below.Those of ordinary skill in the art will appreciate that, unlessotherwise indicated, each embodiment of each individual moiety or groupmay be independently combined with each embodiment of each otherindividual moiety or group in compounds of the present invention.

1. L Group Embodiments

In certain embodiments, L is a bivalent, branched or unbranched,saturated or unsaturated, C₂₋₆ hydrocarbon chain wherein methyleneunit(s) of L is/are are independently replaced by —O—, —S—, —NH—,—C(O)—, —CF₂—, —C(═CH₂)—, —CH═CH—, or an optionally substituted arylene,heteroarylene, C₃-C₆ cycloalkylene, C₃-C₆ heterocycloalkylene, or an8-10-membered bicyclic heterocyclic moiety, and wherein L is optionallysubstituted by one or more groups selected from halogen, C₁-C₆ alkyl,phenyl, —CH₂(phenyl), —CH₂-phenol, —CH(phenyl)₂, —NH₂, —NHC(O)CH₃,—NHC(O)NHCH₂CH₃, —C(O)NH₂, —C(O)NHCH₂CH₃, —CH₂C(O)OCH₂phenyl,—(CH₂)₂SCH₃, —(CH₂)₂C(O)NH₂, —(CH₂)₂C(O)OH, an 8-10 membered bicyclicaryl ring, a 5-6 membered heteroaryl ring having 1-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, an 8-10membered bicyclic heteroaryl ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, a 5- to 7-membered monocyclichaving 1-2 heteroatoms independently selected from nitrogen, oxygen, orsulfur or a 7-10 membered bicyclic heterocyclyl ring having 1-2heteroatoms independently selected from nitrogen, oxygen, or sulfur.

One skilled in the art will appreciate that any number of individualmethylene units within C₂₋₆ hydrocarbon chains may be replaced, asappropriate, by individual moietie(s) according to the presentinvention. One skilled in the art will also appreciate that suchindividual moieties, relative to one another, may be present in anycombination or subcombination within C₂₋₆ hydrocarbon chains.

Exemplary L groups with varied numbers of moieties replacing individualmethylene units within a C₂, C₃, C₄, C₅, or C₆ hydrocarbon chain, andvarious combinations and subcombinations thereof, are described below.

(i) C₂ Hydrocarbon L Groups

In certain embodiments, L is a bivalent, branched or unbranched,saturated or unsaturated, C₂ hydrocarbon chain wherein one or moremethylene units of L is independently replaced by —O—, —S—, —NH—,—C(O)—, —CF₂—, —C(═CH₂)—, —CH═CH—, or an optionally substituted arylene,heteroarylene, C₃-C₆ cycloalkylene, C₃-C₆ heterocycloalkylene, or an8-10-membered bicyclic heterocyclic moiety.

In certain embodiments, L is selected from a bivalent, branched orunbranched, saturated or unsaturated, C₂ hydrocarbon chain wherein oneor more methylene units of L is independently replaced by —NH—, —O—,—C(O)—, —CH═CH—, C₃-C₆ cycloalkylene, C₃-C₆ heterocycloalkylene,8-10-membered bicyclic heterocyclic moiety, an optionally substitutedarylene and optionally substituted heteroarylene, and wherein L isoptionally substituted by one or more groups selected from halogen,substituted or unsubstituted C₁-C₆ alkyl, and —NHC(O)CH₃.

In certain embodiments, L is a bivalent, branched or unbranched,saturated or unsaturated, C₂ hydrocarbon chain wherein one methyleneunit is replaced by —NH—. In certain embodiments, the L group is—NH(CH₃)—. In certain embodiments, L is a C₂ hydrocarbon chain whereinone methylene unit of L is replaced by —NH— and further substituted by—CH₃. In certain embodiments, the L group is —N(CH₃)CH₂—. In certainembodiments, L is a C₂ hydrocarbon chain wherein one methylene unit of Lis substituted. In certain embodiments, a methylene unit of L issubstituted with —NHC(O)CH₃. In certain embodiments, the L group is—CH[NHC(O)CH₃]CH₂—. In certain embodiments, L is a C₂ hydrocarbon chainwherein one methylene unit of L is replaced by —O—. In certainembodiments, the L group is —OCH₂—. In certain embodiments, L is a C₂hydrocarbon chain wherein one methylene unit of L is replaced by —C(O)—.In certain embodiments, the L group is —CH₂C(O)—. In certainembodiments, L is a C₂ hydrocarbon chain wherein one methylene unit of Lis replaced by —O— and one methylene unit of L is replaced by —C(O)—. Incertain embodiments, the L group is —OC(O)—. In certain embodiments, themethylene unit —NH— is substituted. In certain embodiments, the —NH— isoptionally substituted. In certain embodiments, the —NH— is substitutedwith —CH₃, to form —N(CH₃)—. In certain embodiments, L is a C₂hydrocarbon chain wherein one methylene unit of L is replaced by—N(CH₃)—. In certain embodiments, the L group is —CH₂N(CH₃)—. In certainembodiments, L is a C₂ hydrocarbon chain containing a —CH═CH— moiety. Incertain embodiments, L is —CH═CH—. In certain embodiments, L is a C₂hydrocarbon chain wherein one methylene unit of L is replaced by a C₃-C₆cycloalkylene. In certain embodiments, L is a C₂ hydrocarbon chainwherein one methylene unit of L is replaced by a —C(O)— and onemethylene unit is replaced by a C₃-C₆ cycloalkylene. In certainembodiments, the C₃-C₆ cycloalkylene is a C₃ cycloalkylene. In certainembodiments, the L group is

In certain embodiments, the L group is

In certain embodiments, the C₃-C₆ cycloalkylene is a C₄ cycloalkylene.In certain embodiments, the C₃-C₆ cycloalkylene is a C₅ cycloalkylene.In certain embodiments, the C₃-C₆ cycloalkylene is a C₆ cycloalkylene.In certain embodiments, the L group is

In certain embodiments, L is a bivalent, branched or unbranched,saturated or unsaturated, C₂ hydrocarbon chain wherein one methyleneunit is replaced by —C(O)— and one methylene unit is replaced by a C₃-C₆heterocycloalkylene. In certain embodiments, the L group is

In certain embodiments, L is a C₂ hydrocarbon chain wherein onemethylene unit of L is replaced by a 10-membered bicyclic heterocyclicmoiety. In certain embodiments, the 10-membered bicyclic heterocyclicmoiety has one heteroatom. In certain embodiments, the heteroatom isnitrogen. In certain embodiments, the L group is

In certain embodiments, L is a C₂ hydrocarbon chain wherein onemethylene unit of L is replaced by an optionally substituted arylene. Incertain embodiments, the L group is

In certain embodiments, L is a C₂ hydrocarbon chain wherein onemethylene unit of L is replaced by an optionally substituted arylene andone unit of L is replaced by —NH—.In certain embodiments, the L group is

In certain embodiments, L is a C₂ hydrocarbon chain wherein onemethylene unit of L is replaced by an optionally substituted arylene andone unit of L is replaced by —C(O)—. In certain embodiments, the L groupis

In certain embodiments, the arylene is substituted. In certainembodiments, the arylene is a hydroxy-substituted phenylene. In certainembodiments, the L group is

In certain embodiments, L is a C₂ hydrocarbon chain wherein onemethylene unit of L is replaced by an optionally substitutedheteroarylene. In certain embodiments, the heteroarylene is thiophenyl.In certain embodiments, the heteroarylene is furanyl. In certainembodiments, the heteroarylene is indolyl. In certain embodiments, the Lgroup is

In certain embodiments, the L group is

In certain embodiments, the L group is

In certain embodiments, L is a C₂ hydrocarbon chain wherein onemethylene unit of L is replaced by—NH—. In certain embodiments, the L group is —CH₂NH—. In certainembodiments, the L group is —(CH₂)₂NO₂ and no R¹ group is present.

(ii) C₃ Hydrocarbon L Groups

In certain embodiments, L is a bivalent, branched or unbranched,saturated or unsaturated, C₃ hydrocarbon chain wherein one or moremethylene units of L is independently replaced by —O—, —S—, —NH—,—C(O)—, —CF₂—, —C(═CH₂)—, —CH═CH—, or an optionally substituted arylene,heteroarylene, C₃-C₆ cycloalkylene, C₃-C₆ heterocycloalkylene, or an8-10-membered bicyclic heterocyclic moiety.

In certain embodiments, L is selected from a bivalent, branched orunbranched, saturated or unsaturated, C₃ hydrocarbon chain wherein oneor more methylene units of L is independently replaced by —NH—, —O—,—C(O)—, —CF₂—, —C(═CH₂)—, —CH═CH—, C₃-C₆ cycloalkylene, 8-10-memberedbicyclic heterocyclic moiety, an optionally substituted arylene andoptionally substituted heteroarylene, and wherein L is optionallysubstituted by one or more groups selected from halogen, substituted orunsubstituted C₁-C₆ alkyl, -phenyl, —CH(phenyl)₂, —CH₂(phenyl),—NHC(O)CH₃, and NHC(O)NHCH₂CH₃.

In certain embodiments, L is a C₃ hydrocarbon chain wherein onemethylene unit of L is replaced by —C(O)—. In certain embodiments, the Lgroup is —CH₂CH₂C(O)—. In certain embodiments, L is a C₃ hydrocarbonchain wherein one methylene unit of L is replaced by —C(O)— and onemethylene unit is replaced by —NH—. In certain embodiments, the L groupis —C(O)CH₂NH—. In certain embodiments, the L group is —CH₂NHC(O)—. Incertain embodiments, L is a C₃ hydrocarbon chain wherein one methyleneunit of L is replaced by —C(O)—, one methylene unit is replaced by —NH—and one methylene unit is substituted by C₁₋₆ alkyl. In certainembodiments, the L group is —CH[(CH₂)₃CH₃]NHC(O)—. In certainembodiments, L is a C₃ hydrocarbon chain wherein one methylene unit of Lis replaced by —C(O)— and one methylene unit is replaced by —O—. Incertain embodiments, the L group is —CH₂OC(O)—. In certain embodiments,L is a C₃ hydrocarbon chain wherein one methylene unit of L is replacedby —C(O)—, one methylene unit is replaced by —NH— and one methylene isoptionally substituted by —CH₃, —(CH₃)(CH₃) (i.e., dimethyl) or —CH₂CH₃.In certain embodiments, the L group is —C(O)NHC(CH₃)—. In certainembodiments, the L group is —C(O)NHCH(CH₂CH₃)—. In certain embodiments,the L group is

—C(O)NHCH[CH₂CH(CH₃)(CH₃)]— In certain embodiments, L is a C₃hydrocarbon chain wherein one methylene unit of L is replaced by —C(O)—and one or two methylene units are optionally substituted by —CH₃. Incertain embodiments, the L group is

—CH₂CH(CH₃)C(O)—. In certain embodiments, the L group is

—CH(CH₃)CH₂C(O)—. In certain embodiments, the L group is—CH(CH₃)CH(CH₃)C(O)—. In certain embodiments, L is a C₃ hydrocarbonchain wherein one methylene unit of L is replaced by —C(O)— and onemethylene is optionally substituted by —(CH₃)(CH₃) (i.e., dimethyl). Incertain embodiments, the L group is —CH₂C[(CH₃)(CH₃)]C(O)— (i.e., a C₃hydrocarbon chain containing a 3,3-dimethyl substituted methylene). Incertain embodiments, the L group is —C[(CH₃)(CH₃)]CH₂C(O)— (i.e., a C₃hydrocarbon chain containing a 2,2-dimethyl substituted methylene). Incertain embodiments, L is a C₃ hydrocarbon chain wherein one methyleneunit of L is replaced by

—C(O)— and one methylene is optionally substituted by —NHC(O)(CH₃). Incertain embodiments, the L group is —CH₂CH[NHC(O)CH₃]C(O)—. In certainembodiments, L is a C₃ hydrocarbon chain wherein one methylene unit of Lis replaced by —C(O)—, one methylene unit is replaced by —NH— and onemethylene is optionally substituted by —CH(CH₃)₂. In certainembodiments, the L group is —C(O)NHCH[CH(CH₃)(CH₃)]—. In certainembodiments, the L group is —C(O)NHCH₂—. In certain embodiments, L is aC₃ hydrocarbon chain wherein one methylene unit of L is replaced by—C(O)—, one methylene unit is replaced by —O—. In certain embodiments,the L group is —CH₂OC(O)—. In certain embodiments, L is a C₃ hydrocarbonchain wherein one or two methylene units of L are replaced by —C(O)—. Incertain embodiments, the L group is —CH₂CH₂C(O)—. In certainembodiments, the L group is —C(O)CH₂C(O)—. In certain embodiments, L isa C₃ hydrocarbon chain wherein one methylene unit of L is replaced by—NH—. In certain embodiments, the L group is —CH₂CH₂NH—. In certainembodiments, L is a C₃ hydrocarbon chain wherein one methylene unit of Lis replaced by —O—. In certain embodiments, the L group is —CH₂OCH₂—. Incertain embodiments, L is a C₃ hydrocarbon chain wherein one methyleneunit of L is replaced by —C(═CH₂)—. In certain embodiments, L is a C₃hydrocarbon chain wherein one methylene unit of L is replaced by—C(═CH₂)— and one methylene unit is replaced by —C(O)—. One of ordinaryskill in the art will recognize that such a —C(═CH₂)— may exist withinthe hydrocarbon chain backbone or may be “exo” to the backbone chainthus forming and alkylidene group. By way of example, such an L grouphaving a —C═CH₂— within the hydrocarbon chain includes—CH═CHC(O)— or —CH═CHC(O)O—. By way of example, such an L group having asubstituted —C═CH₂— within the hydrocarbon chain includes—CH═C(CH₃)C(O)—, —CH═C(phenyl)-C(O)—. and —CH═CHCF₂. By way of example,such an L group having an alkylidene branched chain includes—CH₂C(═CH₂)C(O)—. In certain embodiments, L is a C₃ hydrocarbon chainwherein one methylene unit of L is replaced by —C(O)— and one methyleneunit is substituted by phenyl. In certain embodiments, the L group is—CH₂CH(phenyl)C(O)—. In certain embodiments, the L group is—CH(phenyl)CH₂C(O)—. In certain embodiments, L is a C₃ hydrocarbon chainwherein one methylene unit of L is replaced by —C(O)— and one methyleneunit is substituted by —NHC(O)NHCH₂CH₃. In certain embodiments, the Lgroup is —CH₂CH[NHC(O)NHCH₂CH₃]C(O)—. In certain embodiments, L is a C₃hydrocarbon chain wherein one methylene unit of L is replaced by —C(O)—,one methylene unit is replaced by —NH— and one methylene unit issubstituted by phenyl or —CH(phenyl)₂. In certain embodiments, the Lgroup is —C(O)NHCH(phenyl)-. In certain embodiments, the L group is—C(O)NHCH[CH(phenyl)₂]—. In certain embodiments, L is a C₃ hydrocarbonchain wherein one methylene unit of L is replaced by —C(O)—, onemethylene unit is replaced by —NH—, and one methylene unit issubstituted by benzyl. In certain embodiments, the L group is—C(O)NHCH[CH₂(phenyl)]—. In certain embodiments, L is a C₃ hydrocarbonchain wherein one methylene unit of L is replaced by —CF₂—. In certainembodiments, the L group is —(CH₂)₂CF₂—. In certain embodiments, L is aC₃ hydrocarbon chain wherein one methylene unit of L is replaced by aC₃-C₆ cycloalkylene. In certain embodiments, the L group is

In certain embodiments, L is a C₃ hydrocarbon chain wherein onemethylene unit of L is replaced by —C(O)—, one methylene unit isreplaced by —NH— and one methylene unit is replaced by a C₃-C₆cycloalkylene. In certain embodiments, the L group is

In certain embodiments, L is a C₃ hydrocarbon chain wherein onemethylene unit of L is replace by —C(O)—, one methylene unit is replacedby —NH— and one methylene unit is further substituted with a C₃-C₆ alkylgroup. In certain embodiments, the C₃-C₆ alkyl group is C₃-C₆cycloalkyl. In certain embodiments, the L group is

In certain embodiments, L is a C₃ hydrocarbon chain wherein onemethylene unit of L is replaced by an optionally substituted arylene. Incertain embodiments, L is a C₃ hydrocarbon chain wherein one methyleneunit of L is replaced by —O— and one methylene unit is replaced by anoptionally substituted arylene. In certain embodiments, the arylene isphenylene. In certain embodiments, the L group is

In certain embodiments, the arylene is a substituted. In certainembodiments, the arylene is a hydroxy-substituted phenylene. In certainembodiments, the L group is

In certain embodiments, L is a C₃ hydrocarbon chain wherein onemethylene unit of L is replaced by an optionally substitutedheteroarylene. In certain embodiments, the heteroarylene is thiophenyl.In certain embodiments, the heteroarylene is furanyl. In certainembodiments, the heteroarylene is indolyl. In certain embodiments, the Lgroup is

In certain embodiments, the L group is

In certain embodiments, the L group is

In certain embodiments, the L group is

(iv) C₄ Hydrocarbon L Groups

In certain embodiments, L is a bivalent, branched or unbranched,saturated or unsaturated, C₄ hydrocarbon chain wherein one or moremethylene units of L is independently replaced by —O—, —S—, —NH—,—C(O)—, —CF₂—, —C(═CH₂)—, —CH═CH—, or an optionally substituted arylene,heteroarylene, C₃-C₆ cycloalkylene, C₃-C₆ heterocycloalkylene, or an8-10-membered bicyclic heterocyclic moiety.

In certain embodiments, L is selected from a bivalent, branched orunbranched, saturated or unsaturated, C₄ hydrocarbon chain wherein oneor more methylene units of L is independently replaced by —NH—, —O—,—C(O)—, —C(═CH₂)—, —CH═CH—, C₃-C₆ cycloalkylene, 8-10-membered bicyclicheterocyclic moiety, an optionally substituted arylene and optionallysubstituted heteroarylene, and wherein L is optionally substituted byone or more groups selected from halogen, and substituted orunsubstituted C₁-C₆ alkyl.

In certain embodiments, L is a bivalent, branched or unbranched,saturated or unsaturated C₄ hydrocarbon chain wherein one or moremethylene units of L is independently replaced by —NH—, —C(O)—, or aC₃-C₆ cycloalkylene. In certain embodiments, L is a C₄ hydrocarbon chainwherein one methylene unit of L is replaced by —C(O)—. In certainembodiments, L is —CH₂CH₂CH₂C(O)—. In certain embodiments, L is—CH(CH₃)CH₂C(O)—. In certain embodiments, L is a C₄ hydrocarbon chainwherein two methylene units of L are replaced by —C(O)—. In certainembodiments, the L group is —C(O)CH₂CH₂C(O)—. In certain embodiments,the L group is —C(O)CH₂CH(CH₃)—. In certain embodiments, the L group is—C(O)CH(CH₃)CH₂—. In certain embodiments, the L group is. In certainembodiments, L is a C₄ hydrocarbon chain wherein one methylene unit of Lis replaced by —C(O)—, one methylene unit is replaced by —NH— and onemethylene unit is substituted by —NH₂. In certain embodiments, the Lgroup is —(CH₂)₂C(O)NH—. In certain embodiments, L is a C₄ hydrocarbonchain wherein one methylene unit of L is replaced by a C₃-C₆cycloalkylene. In certain embodiments, L is

In certain embodiments, L is a C₄ hydrocarbon chain wherein onemethylene unit of L is replaced by an —O—, one methylene unit of L isreplaced by a C₃-C₆ cycloalkylene wherein the C₃-C₆ cycloalkylene isfurther substituted by a C₁-C₆ alkyl group. In certain embodiments, theL group is

In certain embodiments, L is a C₄ hydrocarbon chain wherein onemethylene unit of L is replaced by —C(O)— and wherein L is substitutedby an 8-10 membered bicyclic heteroaryl ring having 1-4 heteroatoms.Exemplary such rings include 1,3-dioxoisoindolinyl. In certainembodiments, the L group is

In certain embodiments, L is a C₄ hydrocarbon chain wherein onemethylene unit of L is replaced by —C(O)— and one methylene unit isreplaced by —NH—. In certain embodiments, L is —NH(CH₂)₂C(O)—. Incertain embodiments, the L group is —C(O)NH(CH₂)₂—. In certainembodiments, the L group is —NHC(O)(CH₂)₂—. In certain embodiments, L isa C₄ hydrocarbon chain wherein one methylene unit of L is replaced by—C(O)—, one methylene unit is replaced by —NH—, and one methylene issubstituted by —OH. In certain embodiments, the L group is—C(O)NHCH[CH₂(OH)]—. In certain embodiments, L is a C₄ hydrocarbon chainwherein one methylene unit of L is replaced by —C(O)—, one methyleneunit is replaced by —NH— and one methylene unit is substituted by —CH₃,—CH₂CH₃, —(CH₂)₃CH₃, —(CH₃)₂, —CH[(CH₃)(CH₃)], —CH₂CH[(CH₃)(CH₃)] or

In certain embodiments, the L group is —CH₂C(O)NHCH(CH₃)—. In certainembodiments, the L group is —CH₂C(O)NHCH[CH(CH₃)(CH₃)]—. In certainembodiments, the L group is —CH₂C(O)NHCH(CH₂CH₃)—. In certainembodiments, the L group is —CH₂C(O)NHCH[CH₂CH(CH₃)(CH₃)]—. In certainembodiments, the L group is

In certain embodiments, L is —CH[(CH₂)₃CH₃]NHC(O)CH₂—. In certainembodiments, L is a C₄ hydrocarbon chain wherein one methylene unit of Lis replaced by —C(O)— and one methylene unit is replaced by —O—. Incertain embodiments, L is —C(O)O(CH₂)₂—. In certain embodiments, the C₄hydrocarbon chain is an alkenylene. In certain embodiments, L is—CH═CHC(O)NH—. In certain embodiments, L is a C₄ hydrocarbon chainwherein one methylene unit of L is replaced by an optionally substitutedarylene. In certain embodiments, the arylene is phenylene. In certainembodiments, the arylene is a substituted. In certain embodiments, thearylene is a substituted phenylene. In

(v) C₅ Hydrocarbon L Groups

In certain embodiments, L is a bivalent, branched or unbranched,saturated or unsaturated, C₅ hydrocarbon chain wherein one or moremethylene units of L is independently replaced by —O—, —S—, —NH—,—C(O)—, —CF₂—, —C(═CH₂)—, —CH═CH—, or an optionally substituted arylene,heteroarylene, C₃-C₆ cycloalkylene, C₃-C₆ heterocycloalkylene, or an8-10-membered bicyclic heterocyclic moiety.

In certain embodiments, L is a bivalent, branched or unbranched,saturated or unsaturated, C₅ hydrocarbon chain wherein one or moremethylene units of L is independently replaced by —NH—, —O—, —C(O)—, andan optionally substituted arylene, and wherein L is optionallysubstituted by one or more groups selected from substituted orunsubstituted C₁-C₆ alkyl, and —CH₂C(O)OH.

In certain embodiments, L is a C₅ hydrocarbon chain wherein onemethylene unit of L is replaced by —C(O)—. In certain embodiments, the Lgroup is

—CH(CH₃)CH(CH₃)C(O)—. In certain embodiments, the L group is—CH₂C[(CH₃)(CH₃)]C(O)—. In certain embodiments, the L group isC[(CH₃)(CH₃)]CH₂C(O)—. In certain embodiments, L is a C₅ hydrocarbonchain wherein one methylene unit of L is replaced by —C(O)— and onemethylene unit is replaced by —NH—. In certain embodiments, L is—C(O)NH(CH₂)₃—. In certain embodiments, L is—(CH₂)₂NHC(O)CH₂—. In certain embodiments, L is a C₅ hydrocarbon chainwherein two methylene units of L are replaced by —C(O)— and onemethylene unit is replaced by —NH—. In certain embodiments, L is—(CH₂)₂C(O)NHNH—. In certain embodiments, L is a C₅ hydrocarbon chainwherein one methylene unit of L is replaced by —O—. In certainembodiments, the L group is OC[(CH₃)(CH₃)]CH₂—. In certain embodiments,L is a C₅ hydrocarbon chain wherein one methylene unit of L is replacedby —C(O)— and one methylene unit is replaced by —O—. In certainembodiments, the L group is —CH₂C(O)OCH₂CH₂— In certain embodiments, Lis a C₅ hydrocarbon chain wherein one methylene unit of L is replaced by—C(O)—, one methylene unit is replaced by —NH—, and one methylene issubstituted by—OH. In certain embodiments, the L group is —C(O)NHCH[CH(CH₃)(OH)]—. Incertain embodiments, L is a C₅ hydrocarbon chain wherein one or twomethylene units of L are replaced by —C(O)—, one methylene unit isreplaced by —NH—, and one methylene unit is substituted with —OH. Incertain embodiments, the L group is —C(O)NHCH[CH₂C(O)OH]—. In certainembodiments, L is a C₅ hydrocarbon chain wherein one methylene unit of Lis replaced by an optionally substituted arylene. In certainembodiments, the arylene is phenylene. In certain embodiments, thearylene is a substituted. In certain embodiments, the arylene is asubstituted phenylene.

(iv) C₆ Hydrocarbon L Groups

In certain embodiments, L is a bivalent, branched or unbranched,saturated or unsaturated, C₆ hydrocarbon chain wherein one or moremethylene units of L is independently replaced by —O—, —S—, —NH—,—C(O)—, —CF₂—, —C(═CH₂)—, —CH═CH—, or an optionally substituted arylene,heteroarylene, C₃-C₆ cycloalkylene, C₃-C₆ heterocycloalkylene, or an8-10-membered bicyclic heterocyclic moiety.

In certain embodiments, L is a bivalent, branched or unbranched,saturated or unsaturated, C₆ hydrocarbon chain wherein one or moremethylene units of L is independently replaced by —NH—, —O—, —C(O)—,—C(═CH₂)—, —CH═CH—, C₃-C₆ cycloalkylene, 8-10-membered bicyclicheterocyclic moiety, an optionally substituted arylene and optionallysubstituted heteroarylene, and wherein L is optionally substituted byone or more groups selected from halogen, substituted or unsubstitutedC₁-C₆ alkyl, —CH₂CH₂C(O)OH,

—(CH₂)₂C(O)NH₂, —C(O)NH₂, —NHC(O)CH₃, —(CH₂)₂SCH₃,—(CH₂)₃NHC(O)NH₂, —(CH₂)₂C(O)OCH₂-phenyl, —NHC(O)NHCH₂CH₃ and a 5- to7-membered monocyclic having 1-2 heteroatoms independently selected fromnitrogen, oxygen, or sulfur

In certain embodiments, L is a C₆ hydrocarbon chain wherein onemethylene unit of L is replaced by —NH—. In certain embodiments, the Lgroup is —CH[CH(CH₃)(CH₂CH₃)]NH—. In certain embodiments, L is a C₆hydrocarbon chain wherein one methylene unit of L is replaced by —C(O)—and one methylene unit is replaced by —O—. In certain embodiments, the Lgroup is —CH₂CH₂C(O)OCH₂CH₂—. In certain embodiments, L is a C₆hydrocarbon chain wherein one or two methylene units of L are replacedby —C(O)—, one methylene unit is replaced by —NH— and one methylene unitis substituted by —OH. In certain embodiments, the L group is—C(O)NHCH[CH₂CH₂C(O)OH]—. In certain embodiments, L is a C₆ hydrocarbonchain wherein one methylene unit of L is replaced by —C(O)— and one ortwo methylene units are replaced by —NH—. In certain embodiments, L is—CH₂C(O)NH(CH₂)₃—. In certain embodiments, L is —CH[(CH₂)₂C(O)NH₂]NH—.In certain embodiments, the L group is C(O)NHCH[CH(CH₃)(CH₃)]—. Incertain embodiments, the L group is C(O)NHCH[CH(CH₃)(CH₃)]—. In certainembodiments, L is a C₆ hydrocarbon chain wherein two methylene units ofL are replaced by —C(O)— and one methylene unit is replaced by —NH— andone methylene unit is substituted by —NH₂ or

—C(O)NHCH₂CH₃. In certain embodiments, the L group is—C(O)NHCH[(CH₂)₂C(O)NH₂]—. In certain embodiments, the L group is—C(O)NHCH[C(O)NH₂](CH₂)₂—. In certain embodiments, the L group is—(CH₂)₂CH[NHC(O)NHCH₂CH₃]C(O)NH—. In certain embodiments, L is a C₆hydrocarbon chain wherein one methylene unit of L is replaced by —C(O)—,one methylene unit is replaced by —NH—, one methylene unit is replacedby —S— which is further substituted by a C₁₋₆ alkyl. In certainembodiments, the L group is —C(O)NHCH[(CH₂)₂SCH₃]—. In certainembodiments, L is a C₆ hydrocarbon chain wherein one methylene unit of Lis replaced by—C(O)—, one methylene unit is replaced by —NH—, and one methylene unitis replaced by —O—. In certain embodiments, the L group is—NHCH₂C(O)OCH₂CH₂—. In certain embodiments, L is a C₆ hydrocarbon chainwherein one methylene unit of L is replaced by —C(O)— and one methyleneunit is replaced by —NH—, and one methylene unit is further substitutedby a C₁₋₆ alkyl. In certain embodiments, the L group is—C(O)NHCH[CH₂CH(CH₃)₂]—. In certain embodiments, the L group is—C(O)NHCH[CH(CH₃)(CH₂CH₃)]. In certain embodiments, the L group is—C(O)NHCH[CH(CH₃)(CH₂CH₃)]—. In certain embodiments, L is a C₆hydrocarbon chain wherein one or two methylene units of L are replacedby —C(O)— and one methylene unit is replaced by —NH—. In certainembodiments, the L group is—CH₂CH[NHC(O)CH₃]C(O)—. In certain embodiments, the L group is—CH₂CH[NHC(O)CH₃]C(O)—. In certain embodiments, L is a C₆ hydrocarbonchain wherein one or two methylene units of L are replaced by —C(O)—,one methylene unit is replaced by—NH—, one methylene unit is replaced by —O— and one methylene unit issubstituted with —OCH₂-phenyl. In certain embodiments, the L group is—C(O)NH—CH[(CH₂)₂C(O)OCH₂-phenyl]-. In certain embodiments, L is a C₆hydrocarbon chain wherein one or two methylene units of L are replacedby —NH— and one methylene unit is substituted with —C(O)NH₂. In certainembodiments, the L group is —CH[(CH₂)₃NHC(O)NH₂]NH—. In certainembodiments, the L group is —(CH₂)₂CH[C(O)NH₂]NH—. In certainembodiments, L is a C₆ hydrocarbon chain wherein one methylene unit of Lis replaced by —NH—, one methylene unit is replaced by —C(O)— and onemethylene unit is substituted with —C(O)NH₂. In certain embodiments, Lis a C₆ hydrocarbon chain wherein two methylene units of L are replacedby —C(O)—, one methylene unit is replaced by —NH—, one methylene unit isreplaced by —O— and one methylene substituted with —NHCH₂CH₃. In certainembodiments, the L group is—CH₂CH[NHC(O)NHCH₂CH₃]C(O)O—. In certain embodiments, L is a C₆hydrocarbon chain wherein one methylene unit of L is replaced by anoptionally substituted arylene. In certain embodiments, the arylene isphenylene. In certain embodiments, the arylene is a substituted. Incertain embodiments, the arylene is a substituted phenylene. In certainembodiments, L is a C₆ hydrocarbon chain wherein one methylene unit of Lis replaced by —NH—, one methylene unit is replaced by —C(O)—, onemethylene unit is substituted by a morpholine ring and one methyleneunit is further substituted by a —CH₃ group. In certain embodiments, theL group is —C(O)[morpholino]NHCH[CH(CH₃)(CH₂)]—. In certain embodiments,it will further be appreciated that the -L-R¹ moiety of a compound ofFormulae I, and/or I′ is —C(O)[morpholino]NHCH[CH(CH₃)(CH₂CH₃)]—,wherein R¹ is a —CH₃.

In summary, a list of Exemplary L groups include —NHCH₂—, —N(CH₃)—,—CH₂CH₂C(O)—, —CH═CHC(O)—, —CH═CHC(O)O—, —NHCH₂C(O)—, —NH(CH₂)₂C(O)—,—CH₂C(═CH₂)C(O)—, —CH₂CH₂CH₂C(O)—,

—NHCH₂CH₂C(O)—, —CH₂CH₂C(O)OCH₂CH₂—, —C(O)CH₂C(O)—, —C(O)CH₂CH₂C(O)—,—NH(CH₃)—, —N(CH₃)CH₂—, —CH[NHC(O)CH₃]CH₂—, —OCH₂—, —CH₂C(O)—, —OC(O)—,—N(CH₃)—, —N(CH₃)—. —CH₂N(CH₃)—, —CH═CH—,

—CH₂NH—, —(CH₂)₂NO₂, —CH₂CH₂C(O)—, —C(O)CH₂NH—, —CH₂NHC(O)—,—CH[(CH₂)₃CH₃]NHC(O)—, —CH₂OC(O)—, —C(O)NHC(CH₃)—, —C(O)NHCH(CH₂CH₃)—,—C(O)NHCH[CH₂CH(CH₃)(CH₃)]—, —CH₂CH(CH₃)C(O)—, —CH(CH₃)CH₂C(O)—,—CH(CH₃)CH(CH₃)C(O)—, —CH₂C[(CH₃)(CH₃)]C(O)—, —C[(CH₃)(CH₃)]CH₂C(O)—,—NHC(O)(CH₃), —CH₂CH[NHC(O)CH₃]C(O)—, —C(O)NHCH[CH(CH₃)(CH₃)]—,—C(O)NHCH₂—, —CH₂OC(O)—, —CH₂CH₂C(O)—, —C(O)CH₂C(O)—, —CH₂CH₂NH—,—CH₂OCH₂—, —CH═CHC(O)—, or —CH═CHC(O)O—, —CH═C(CH₃)C(O)—,—CH═C(phenyl)-C(O)—, —CH═CHCF₂, —CH₂C(═CH₂)C(O)—, —CH₂CH(phenyl)C(O)—,—CH(phenyl)CH₂C(O)—, —NHC(O)NHCH₂CH₃, —CH₂CH[NHC(O)NHCH₂CH₃]C(O)—,—C(O)NHCH(phenyl)-, —C(O)NHCH[CH(phenyl)₂]—, —C(O)NHCH[CH₂(phenyl)]-,—(CH₂)₂CF₂—, C₃-C₆ cycloalkylene,

—CH₂CH₂CH₂C(O)—, —CH(CH₃)CH₂C(O)—, —C(O)CH₂CH₂C(O)—, —C(O)CH₂CH(CH₃)—,—C(O)CH(CH₃)CH₂—, —(CH₂)₂C(O)NH—,

1,3-dioxoisoindolinyl,

—NH(CH₂)₂C(O)—, —C(O)NH(CH₂)₂—., —NHC(O)(CH₂)₂—, —C(O)NHCH[CH₂(OH)]—,—CH₂C(O)NHCH(CH₃)—, —CH₂C(O)NHCH[CH(CH₃)(CH₃)]—, —CH₂C(O)NHCH(CH₂CH₃)—,—CH₂C(O)NHCH[CH₂CH(CH₃)(CH₃)]—,

—CH[(CH₂)₃CH₃]NHC(O)CH₂—, —C(O)O(CH₂)₂—, —CH═CHC(O)NH—,—CH(CH₃)CH(CH₃)C(O)—, —CH₂C[—(CH₃)(CH₃)]C(O)—, —C[—(CH₃)(CH₃)]CH₂C(O)—,—C(O)NH(CH₂)₃—, —(CH₂)₂NHC(O)CH₂—, —(CH₂)₂C(O)NHNH—,I—OC[—(CH₃)(CH₃)]CH₂—, —CH₂C(O)OCH₂CH₂—, —C(O)NHCH[CH(CH₃)(OH)]—,—C(O)NHCH[CH₂C(O)OH]—, —CH[CH(CH₃)(CH₂CH₃)]NH—, —CH₂CH₂C(O)OCH₂CH₂,—C(O)NHCH[CH₂CH₂C(O)OH]—, —CH₂C(O)NH(CH₂)₃—, —CH[(CH₂)₂C(O)NH₂]NH—,—C(O)NHCH[CH(CH₃)(CH₃)]—, —C(O)NHCH[CH(CH₃)(CH₃)]—,—C(O)NHCH[(CH₂)₂C(O)NH₂]—, —C(O)NHCH[C(O)NH₂](CH₂)₂—,—(CH₂)₂CH[NHC(O)NHCH₂CH₃]C(O)NH—, —C(O)NHCH[(CH₂)₂SCH₃]—,—NHCH₂C(O)OCH₂CH₂—, —C(O)NHCH[CH₂CH(CH₃)₂]—,—C(O)NHCH[CH(CH₃)(CH₂CH₃)]—.—C(O)NHCH[CH(CH₃)(CH₂CH₃)]—,—CH₂CH[NHC(O)CH₃]C(O)—, —CH₂CH[NHC(O)CH₃]C(O)—,—C(O)NH—CH[(CH₂)₂C(O)OCH₂-phenyl], —CH[(CH₂)₃NHC(O)NH₂]NH—,—(CH₂)₂CH[C(O)NH₂]NH—, —(CH₂)₂CH[C(O)NH₂]NHC(O)CH₃—,—CH₂CH[NHC(O)NHCH₂CH₃]C(O)O— and —C(O)[morpholino]NHCH[CH(CH₃)(CH₂)]—.

ii. R¹ Group Embodiments

As defined generally above, the R¹ group of Formulae I, and/or I′ is R¹is hydrogen, F, CF₃, C₁-C₄ alkyl, —OH, —C(O)CH₃, —NH(OR), —NR₂, —NHNR₂,—SO₂R, —NH-phenyl, —SO₂-phenyl, -phenyl-NO₂, or —OR, wherein each R isindependently hydrogen, oxygen, or an optionally substituted groupselected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic.

Exemplary R¹ groups include: hydrogen, —F, —CF₃, —CH₃, —OH, —C(O)CH₃,—C(O)CF₃, —NH₂, —NH₂NH₂, —NHCH₂CH₃, —SO₂-methyl,

wherein each R is independently hydrogen or an optionally substitutedgroup selected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic.

In certain embodiments, R¹ is hydrogen. In certain embodiments, R¹ is F.In certain embodiments, R¹ is CF₃. In certain embodiments, R¹ is C₁-C₄alkyl. In certain embodiments, R¹ is —CH₃. In certain embodiments, R¹ ismethyl. In certain embodiments, R¹ is —OH. In certain embodiments, R¹ is—C(O)CH₃. In certain embodiments, R¹ is —C(O)CF₃. In certainembodiments, R¹ is —NH₂. In certain embodiments, R¹ is —NH₂NH₂. Incertain embodiments, R¹ is —NHCH₂CH₃. In certain embodiments, R¹ is—SO₂-methyl. In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is

In certain embodiments, R¹ is —OR. In certain embodiments, R¹ is —OR,wherein R is an optionally substituted C₁₋₆ aliphatic. In certainembodiments, R¹ is —OCH₂CH₃. In certain embodiments, R¹ is —NHR, whereinR is as defined herein. In certain embodiments, R¹ is —NH(OR), wherein Ris as defined herein. In certain embodiments, R¹ is —ONH₂. In certainembodiments, R¹ is —NR₂, wherein R is as defined herein.

iii. R² Group Embodiments

As defined generally above, the R² group of Formulae I and/or I′ is R²is —C(O)X, wherein X is independently R, —C(O)NHNH₂, —OR, a hydrogen,aryloxy, amino, alkylamino, dialkylamino, heteroaryloxy, hydrazine, a6-10 membered aryl ring, a 5-6 membered heteroaryl ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur,wherein each R is independently hydrogen or an optionally substitutedgroup selected from C₁₋₆ aliphatic or C₁₋₆ heteroaliphatic. In certainembodiments, R² is —C(O)X, wherein X is selected from R, —OR, hydrazine,or a hydrogen. In certain embodiments, R² is —C(O)X. In certainembodiments, R² is —C(O)H. In certain embodiments, R² is —C(O)OH. Incertain embodiments, R² is —C(O)OR. In certain embodiments, R² is—C(O)NHNH₂.

iv. R³ Group Embodiments

As defined generally above, the R³ group of Formulae I and/or I′ is asubstituted or unsubstituted, branched or unbranched, saturated orunsaturated, C₁₀-C₂₅ aliphatic. In certain embodiments, R³ is asubstituted or unsubstituted, branched or unbranched C₁₀-C₁₅ aliphatic.In certain embodiments, R³ is a substituted or unsubstituted, branchedor unbranched C₁₀-C₁₅ alkenyl. In certain embodiments, R³ is asubstituted or unsubstituted, branched or unbranched C₁₀-C₁₂ aliphatic.In certain embodiments, R³ is a substituted or unsubstituted, branchedor unbranched C₁₂ aliphatic. In certain embodiments, R³ is anunsubstituted, branched C₁₂ aliphatic. In certain embodiments, R³ is asubstituted, branched C₁₂ aliphatic. In certain embodiments, R³ is abranched C₁₂ alkenyl group. In certain embodiments, R³ is—CH₂CH═C(CH₃)CH₂CH₂CH═C(CH₃)CH₂CH₂CH═C(CH₃)(CH₃). In certainembodiments, R³ is a substituted, branched C₁₅ aliphatic. In certainembodiments, R³ is a branched C₁₅ alkenyl group. In certain embodiments,R³ is —CH₂CH═C(CH₃)CH₂CH₂CH═C(CH₃)CH₂CH₂CH═C(CH₃)(CH₃). In certainembodiments, R³ is a substituted, branched C₁₆ aliphatic. In certainembodiments, R³ is a branched C₁₆ alkenyl group. In certain embodiments,R³ is —CH₂CH═C(CH₃)CH₂CH₂CH₂CH(CH₃)CH₂CH₂CH₂CH(CH₃)CH₂CH₂CH₂CH(CH₃)(CH₃). In certain embodiments, R³ is asubstituted, branched C₂₀ aliphatic. In certain embodiments, R³ is abranched C₂₀ alkenyl group. In certain embodiments, R³ is—CH₂CH═C(CH₃)CH₂CH₂CH₂CH(CH₃) CH₂CH₂CH₂CH(CH₃)CH₂CH₂CH₂CH(CH₃)(CH₃).

v. Y Group Embodiments

As defined generally above, the Y group is —O—, —N—, —S—, —Se—, —S(O)—,—S(═N)—, —S(O)₂—, —Se(O)—, —Se(O)₂—, or —C(═S)—. In certain embodiments,Y is —S—. In certain embodiments, Y is —O—. In certain embodiments, Y is—N—. In certain embodiments, Y is —Se—. In certain embodiments, Y is—S(O)—. In certain embodiments, Y is —S(═N)—. In certain embodiments, Yis —S(O)₂—. In certain embodiments, Y is —Se(O)—. In certainembodiments, Y is —Se(O)₂—.

6. Stereochemistry Embodiments

As described herein, compounds may comprise one or more chiral centers,and thus can exist in various stereoisomeric forms, e.g., enantiomers,diastereomers, or geometric isomers). Thus, inventive compounds andpharmaceutical compositions thereof may be in the form of a racemiccompound, an individual enantiomer (e.g., enantiomerically pure), anindividual diastereomer (e.g., diastereomerically pure), an individualgeometric isomer (e.g., geometrically pure), or may be in the form of amixture of stereoisomers. In certain embodiments, compounds of thepresent invention are racemic compounds. In certain embodiments,compounds of the present invention are enantioenriched compounds. Incertain embodiments, compounds of the present invention arediasteriomerically enriched compounds. In certain embodiments, whereinone or more double bonds is present, compounds of the present inventionmay be geometrically enriched compounds. In certain embodiments,compounds of the present invention are provided such that 75% of thepreparation is of the same enantiomer or diastereomer. In certainembodiments, compounds of the present invention are provided such thatat least 80%, 90%, 95%, or 97.5% of the preparation is of the sameenantiomer or diastereomer. In certain embodiments, compounds of thepresent invention are provided such the preparation consists of a singleenantiomer or diastereomer to the limits of detection (i.e.,“enantiopure”).

It will be apparent to one skilled in the art that each chiral center ina provided compound can be present in an (R)-configuration or in an(S)-configuration. In addition, where stereoisomeric forms of providedcompounds may exist, such forms may be present in any ratio relative toone another. One skilled in the art will further understand that ratiosof stereoisomers may vary according to methods by which such compoundsare prepared. Exemplary ratios provided herein are meant to illustratethe present invention, and are not meant to limit the present invention.

With respect to geometric isomerism, the present invention contemplatesboth E and Z isomers wherein there exists one or more double bonds,unless otherwise indicated. In some embodiments, the inventionencompasses compounds as a single geometric isomer substantially free ofother geometric isomers and alternatively, as mixtures of variousisomers, e.g., racemic mixtures of E and Z isomers. In addition to theabove-mentioned compounds per se, this invention also encompassespharmaceutically acceptable derivatives of these compounds andcompositions comprising one or more compounds of the invention and oneor more pharmaceutically acceptable excipients or additives.

Where a stereoisomer is preferred, it may, in some embodiments beprovided substantially free of other stereoisomers, as defined herein.According to certain embodiments, the present invention providescompounds of Formulae I, I′, and/or Ia substantially free of otherstereoisomers.

Enantiomeric and stereoisomeric mixtures may be resolved into theircomponent enantiomers or stereoisomers by well known methods, such aschiral-phase gas chromatography, chiral-phase high performance liquidchromatography, crystallizing a compound as a chiral salt complex, orcrystallizing a compound in a chiral solvent or by enzymatic resolutionof a compound, its precursor or its derivative. Enantiomers andstereoisomers may also be obtained from stereomerically orenantiomerically pure intermediates, reagents, and catalysts bywell-known asymmetric synthetic methods.

Additionally, unless otherwise stated, the present invention encompassescompounds that differ from those explicitly depicted herein only in thepresence of one or more isotopically enriched atoms. For example,compounds having the present structures including the replacement ofhydrogen by deuterium or tritium, or the replacement of a carbon by a¹³C- or ¹⁴C-enriched carbon are within the scope of this invention. Suchcompounds are useful, for example, as analytical tools, as probes inbiological assays, or as therapeutic agents in accordance with thepresent invention. In certain embodiments, the R¹ group of I, I′, and/orIa comprises one or more deuterium atoms. In certain embodiments, the R²group of I, I′, and/or Ia comprises one or more deuterium atoms. Incertain embodiments, the R³ group of I, I′, and/or Ia comprises one ormore deuterium atoms. Mixtures of isomeric forms may be separated and/orpurified by techniques as would be known to one skilled in this art,including but not limited to column chromatography.

As described generally above, the present invention provides a compoundof Formulae I, I′, and/or Ia, having a stereochemistry as depicted inFormula 1a and/or 1b:

or a pharmaceutically, acceptable salt thereof, wherein each variable isdefined above and in classes and subclasses described above and herein.

It will be appreciated that for each racemic compound disclosed herein,individual enantiomers are also contemplated. For example, one of skillin the art would understand that compound N-54 as depicted below:

also contemplates each of its enantiomers:

I. L Group Stereochemistry and R³ Group Stereochemistry

Exemplary L group and R³ group stereochemistry of the present inventionare described below. It will be appreciated that all combinations ofembodiments, as described herein, are contemplated. In some embodiments,the present invention provides a compound having any combination of oneof the L groups and one of the R³ group described below. It will furtherbe appreciated that wherein a specific L group or R³ group is describedgenerally without specifying stereochemistry, the present inventioncontemplates all embodiments of stereochemistry associated with thatgroup.

A. L Group Stereochemistry General Definition of L Group

As generally described above and herein, L is a bivalent, branched orunbranched, saturated or unsaturated, C₂-C₆ hydrocarbon chain whereinone or more methylene units of L is independently replaced by —O—, —S—,—NH—, —C(O)—, —CF₂—, —C(═CH₂)—, —CH═CH—, or an optionally substitutedarylene, heteroarylene, C₃-C₆ cycloalkylene, C₃-C₆ heterocycloalkylene,or an 8-10-membered bicyclic heterocyclic moiety, and wherein L isoptionally substituted by one or more groups selected from halogen,C₁-C₆ alkyl, phenyl, biphenyl, -benzyl, —CH₂-phenol, —CH(phenyl)₂, —OH,—NH₂, —NHC(O)CH₃, —NHC(O)NHCH₂CH₃, —C(O)NH₂,

—C(O)NHCH₂CH₃, —CH₂C(O)OCH₂phenyl, —(CH₂)₂SCH₃, —(CH₂)₂C(O)NH₂,—(CH₂)₂C(O)OH, an 8-10 membered bicyclic aryl ring, a 5-6 memberedheteroaryl ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, an 8-10 membered bicyclic heteroaryl ringhaving 1-4 heteroatoms independently selected from nitrogen, oxygen, orsulfur, a 5- to 7-membered monocyclic having 1-2 heteroatomsindependently selected from nitrogen, oxygen, or sulfur or a 7-10membered bicyclic heterocyclyl ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur.

One Chiral Center (i.e., No Chiral Centers in L)

In some embodiments, a compound of Formula I or Formula I′, havingstereochemistry as depicted in Formula 1a or 1b, contains no chiralcenters in the C₂₋₆ hydrocarbon chain of L. Exemplary such compoundsinclude, for instance, Compound A [(R)-enantiomer; Example 2] andcorresponding (S)-enantiomer.

Two Chiral Centers (i.e., One Chiral Center in L)

In some embodiments, a compound of Formula I or Formula I′, havingstereochemistry as depicted in Formula 1a or 1b, contains one chiralcenter in the C₂₋₆ hydrocarbon chain of L. In certain embodiments, achiral center in the C₂₋₆ hydrocarbon chain of L is at C₂. In certainembodiments, a chiral center in the C₂₋₆ hydrocarbon chain of L is atC₃. In certain embodiments, a chiral center in the C₂₋₆ hydrocarbonchain of L is at C₄. In certain embodiments, a chiral center in the C₂₋₆hydrocarbon chain of L is at C₅. In certain embodiments, a chiral centerin the C₂₋₆ hydrocarbon chain of L is at C₆. One of skill in the artwould recognize that a chiral center in the C₂₋₆ hydrocarbon chain of Lmay be present in either the (R) or (S) configuration. In certainembodiments, a chiral center in the C₂₋₆ hydrocarbon chain of L isenantiopure or enantioenriched in the (R) configuration. In certainembodiments, a chiral center in the C₂₋₆ hydrocarbon chain of L isenantiopure or enantioenriched in the (S) configuration. In certainembodiments, a chiral center in the C₂₋₆ hydrocarbon chain of L ispresent in approximately a 1:1 molar ratio of (R) to (S).

Exemplary stereochemistry present within an L group containing onechiral center in the C₂₋₆ hydrocarbon chain of a compound of Formula Ior Formula I′, and having stereochemistry as depicted in Formula 1a or1b, are as depicted below in Formulae 1l-(i), 1l-(ii), 1l-(iii),1l-(iv), 1l-(v) and/or 1l-(vi):

Exemplary compounds having stereochemistry as depicted in Formulae1l-(v) and 1l-(vi) above for the L group, include: Compound C-2 (Example5b), Compound N-55 (Example 9), Compound N-57 (Example 11), CompoundN-58 (Example 12), Compound N-8 (Example 14), Compound N-3 (Example 15),Compound N-5 (Example 17), Compound N-9 (Example 20), Compound N-12(Example 21), Compound N-60 (Example 23), Compound N-50 (Example 24),Compound N-63 (Example 29), Compound N-66 (Example 34), Compound N-71(Example 46), Compound N-91, and Compound N-92.

In certain embodiments, compounds of the present invention, havingstereochemistry as depicted in Formula 1a, are provided such thatcompounds containing an L group of Formula 1l-(i) and compoundscontaining an L group of Formula 1l-(ii) are present in a 1:1 molarratio. Exemplary such compounds include Compound C (Example 5 andExample 5a), Compound N-2, Compound N-18, Compound N-31 (Example 62),Compound N-34 (Example 41a), Compound N-37, Compound N-40 (Example 32),Compound N-41 (Example 33), Compound N-46 (Example 35), Compound N-47,Compound N-61 (Example 27), Compound N-64 (Example 30), Compound N-65(Example 31), Compound N-77 (Example 65), Compound N-89, Compound N-93,and Compound N-94.

In certain embodiments, compounds of the present invention, havingstereochemistry as depicted in Formula 1b, are provided such thatcompounds containing an L group of Formula 1l-(i) and compoundscontaining an L group of Formula 1l-(ii) are present in a 1:1 molarratio.

In certain embodiments, compounds of the present invention, havingstereochemistry as depicted in Formula 1a, are provided such thatcompounds containing an L group of Formula 1l-(iii) and compoundscontaining an L group of Formula 1l-(iv) are present in a 1:1 molarratio. Exemplary such compounds include Compound N-36; Compound N-78(Example 66); and Compound N-32 (Example 67).

In certain embodiments, compounds of the present invention, havingstereochemistry as depicted in Formula 1b, are provided such thatcompounds containing an L group of Formula 1l-(iii) and compoundscontaining an L group of Formula 1l-(iv) are present in a 1:1 molarratio.

In certain embodiments, compounds of the present invention, havingstereochemistry as depicted in Formula 1a, are provided such thatcompounds containing an L group of Formula 1l-(v) and compoundscontaining an L group of Formula 1l-(vi) are present in a 1:1 molarratio. Exemplary compounds of this type include Compound N-88 andCompound N-95.

In certain embodiments, compounds of the present invention, havingstereochemistry as depicted in Formula 1b, are provided such thatcompounds containing an L group of Formula 1l-(v) and compoundscontaining an L group of Formula 1l-(vi) are present in a 1:1 molarratio.

Three Chiral Centers (i.e., Two Chiral Center in L)

In certain embodiments, a compound of Formula I or Formula I′, havingstereochemistry as depicted in Formula 1a or 1b, contains two chiralcenters in the C₂₋₆ hydrocarbon chain of L. In certain embodiments, twochiral centers in the C₂₋₆ hydrocarbon chain of L are at C₁ and C₂. Incertain embodiments, two chiral centers in the C₂₋₆ hydrocarbon chain ofL are at C₁ and C₃. In certain embodiments, two chiral centers in theC₂₋₆ hydrocarbon chain of L are at C₁ and C₄. In certain embodiments,two chiral centers in the C₂₋₆ hydrocarbon chain of L are at C₁ and C₅.In certain embodiments, two chiral centers in the C₂₋₆ hydrocarbon chainof L are at C₁ and C₆. In certain embodiments, two chiral centers in theC₂₋₆ hydrocarbon chain of L are at C₂ and C₃. In certain embodiments,two chiral centers in the C₂₋₆ hydrocarbon chain of L are at C₂ and C₄.In certain embodiments, two chiral centers in the C₂₋₆ hydrocarbon chainof L are at C₂ and C₅. In certain embodiments, two chiral centers in theC₂₋₆ hydrocarbon chain of L are at C₂ and C₆. In certain embodiments,two chiral centers in the C₂₋₆ hydrocarbon chain of L are at C₃ and C₄.In certain embodiments, two chiral centers in the C₂₋₆ hydrocarbon chainof L are at C₃ and C₅. In certain embodiments, two chiral centers in theC₂₋₆ hydrocarbon chain of L are at C₃ and C₆. In certain embodiments,two chiral centers in the C₂₋₆ hydrocarbon chain of L are at C₄ and C₅.In certain embodiments, two chiral centers in the C₂₋₆ hydrocarbon chainof L are at C₄ and C₆. In certain embodiments, two chiral centers in theC₂₋₆ hydrocarbon chain of L are at C₅ and C₆.

Exemplary stereochemistry present within an L group containing twochiral centers in the C₂₋₆ hydrocarbon chain of a compound of Formula Ior Formula I′, and having a stereochemistry as depicted in Formula 1a or1b, are as depicted below in Formulae 2l-(i), 2l-(ii), 2l-(iii), and2l-(iv):

One of skill in the art would recognize that either of the two chiralcenters in the C₂₋₆ hydrocarbon chain of L may be present in an (R) or(S) configuration. In certain embodiments, both chiral centers in theC₂₋₆ hydrocarbon chain of L are in an (R) configuration. In certainembodiments, both chiral centers in the C₂₋₆ hydrocarbon chain of L arein an (S) configuration. In certain embodiments, one chiral center inthe C₂₋₆ hydrocarbon chain of L is present in an (R) configuration and asecond chiral center in the C₂₋₆ hydrocarbon chain of L is present in an(S) configuration.

In certain embodiments, compounds of Formula I and/or Formula I′, havingstereochemistry as depicted in Formula 1a and/or 1b, are provided suchthat at least one of two chiral centers in the C₂₋₆ hydrocarbon chain ofL is enantiopure or enantioenriched in an (R) or (S) configuration.

In certain embodiments, compounds of Formula I and/or Formula I′, havingstereochemistry as depicted in Formula 1a and/or 1b, are provided suchthat both chiral centers in the C₂₋₆ hydrocarbon chain of L areindependently enantiopure or enantioenriched in an (R) or (S)configuration.

In certain embodiments, compounds of Formula I and/or Formula I′, havingstereochemistry as depicted in Formula 1a and/or 1b, are provided suchthat one of the two chiral centers in the C₂₋₆ hydrocarbon chain of L isenantiopure or enantioenriched in an (R) or (S) configuration, while theother chiral center in the C₂₋₆ hydrocarbon chain of L is present as aracemate.

In certain embodiments, compounds of Formula I and/or Formula I′, havingstereochemistry as depicted in Formula 1a and/or 1b, are provided suchthat both chiral centers in the C₂₋₆ hydrocarbon chain of L are presentas racemates.

In some embodiments, wherein compounds of the present invention areprovided as a mixture of one or more stereoisomers, all possiblestereoisomers of L are present. In some embodiments, wherein compoundsof the present invention are provided as a mixture of stereoisomers, amixture may contain two stereoisomers present in a ratio of about 20:1,18:1, 16:1, 14:1, 12:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, or1:1.

B. R³ Group Stereochemistry

As described generally above and herein, R³ is a substituted orunsubstituted, branched or unbranched, saturated or unsaturated C₁₀-C₂₅aliphatic.

In some embodiments, R³ of Formula I or Formula I′, havingstereochemistry as depicted in Formula 1a or 1b, is of Formula 1r:

In some embodiments, R³ of Formula I or Formula I′, havingstereochemistry as depicted in Formula 1a or 1b, is of general Formula2r:

In certain embodiments, R³ of Formula I or Formula I′, having astereochemistry as depicted in Formula 1a or 1b, is of Formula 2r,wherein 2r is of any of Formulae 2r-(i), 2r-(ii), 2r-(iii), or 2r-(iv):

In certain embodiments, compounds of Formula I and/or Formula I′, havingstereochemistry as depicted in Formula 1a and/or 1b, are provided suchthat at least one of two chiral centers in R³ is enantiopure orenantioenriched in an (R) or (S) configuration.

In certain embodiments, compounds of Formula I and/or Formula I′, havingstereochemistry as depicted in Formula 1a and/or 1b, are provided suchthat two chiral centers in R³ are independently enantiopure orenantioenriched in an (R) or (S) configuration.

In certain embodiments, compounds of Formula I and/or Formula I′, havingstereochemistry as depicted in Formula 1a and/or 1b, are provided suchthat a first chiral center in R³ is enantiopure or enantioenriched in an(R) or (S) configuration, while a second chiral center in R³ is presentas the racemate.

In certain embodiments, compounds of Formula I and/or Formula I′, havingstereochemistry as depicted in Formula 1a and/or 1b, are provided suchthat two chiral centers in R³ are present as racemates.

In some embodiments, wherein R³ is of Formula 2r depicted above,compounds of Formula I and/or Formula I′, having stereochemistry asdepicted in Formula 1a and/or 1b, are provided such that all possiblestereoisomers of Formula 2r are present, three stereoisomers of formula2r are present, two stereoisomers of Formula 2r are present, or onestereoisomer of Formula 2r is present.

In certain embodiments, wherein R³ is of Formula 2r depicted above,compounds of Formula I and/or Formula I′, having stereochemistry asdepicted in Formula 1a and/or 1b, are provided such that only compoundscontaining stereochemistry as depicted in 2r-(i) and 2r-(ii) arepresent.

In certain embodiments, wherein R³ is of Formula 2r depicted above,compounds of Formula I and/or Formula I′, having stereochemistry asdepicted in Formula 1a and/or 1b, are provided such that only compoundscontaining stereochemistry as depicted in 2r-(iii) and 2r-(iv) arepresent.

In certain embodiments, wherein R³ is of Formula 2r depicted above,compounds of Formula I and/or Formula I′, having stereochemistry asdepicted in Formula 1a and/or 1b, are provided such that compoundscontaining stereochemistry as depicted in 2r-(i) and 2r-(ii) are presentin a 1:1 ratio. In certain embodiments, wherein R³ is of Formula 2rdepicted above, compounds of Formula I and/or Formula I′, havingstereochemistry as depicted in Formula 1a and/or 1b, are provided suchthat compounds containing stereochemistry as depicted in 2r-(i) and2r-(ii) are not present in a 1:1 ratio.

In certain embodiments, wherein R³ is of Formula 2r depicted above,compounds of Formula I and/or Formula I′, having stereochemistry asdepicted in Formula 1a and/or 1b, are provided such that compoundscontaining stereochemistry as depicted in 2r-(iii) and 2r-(iv) arepresent in a 1:1 ratio. In certain embodiments, wherein R³ is of Formula2r depicted above, compounds of Formula I and/or Formula I′, havingstereochemistry as depicted in Formula 1a and/or 1b, are provided suchthat compounds containing stereochemistry as depicted in 2r-(iii) and2r-(iv) are not present in a 1:1 ratio.

In certain embodiments, wherein R³ is of Formula 2r depicted above,compounds of Formula I and/or Formula I′, having stereochemistry asdepicted in Formula 1a and/or 1b, are provided such that compoundscontaining stereochemistry as depicted in 2r-(i) and 2r-(ii) are presentin a 1:1 ratio and compounds containing stereochemistry as depicted in2r-(iii) and 2r-(iv) are present in a 1:1 ratio. In certain embodiments,wherein R³ is of Formula 2r depicted above, compounds of Formula Iand/or Formula I′, having stereochemistry as depicted in Formula 1aand/or 1b, are provided such that compounds containing stereochemistryas depicted in 2r-(i) and 2r-(ii) are present in a 1:1 ratio andcompounds containing stereochemistry as depicted in 2r-(iii) and 2r-(iv)are not present in a 1:1 ratio. In certain embodiments, wherein R³ is ofFormula 2r depicted above, compounds of Formula I and/or Formula I′,having stereochemistry as depicted in Formula 1a and/or 1b, are providedsuch that compounds containing stereochemistry as depicted in 2r-(i) and2r-(ii) are not present in a 1:1 ratio and compounds containingstereochemistry as depicted in 2r-(iii) and 2r-(iv) are present in a 1:1ratio.

In certain embodiments, wherein R³ is of Formula 2r depicted above, andwherein compounds of the invention are provided such that stereochemicalconfigurations depicted in each of 2r-(i), 2r-(ii), 2r-(iii), and2r-(iv) are present, the ratio of the sum of compounds containingstereochemistry as depicted in 2r-(i) and 2r-(ii) [i.e., the totalamount of compounds wherein R³ is present in either an (R,R) or (S,S)“cis” configuration] to the sum of compounds containing stereochemistryas depicted in 2r-(iii) and 2r-(iv) [i.e., the total amount of compoundswherein R³ is present in either an (R,S) or (S,R) “trans” configuration]is about 20:1, 18:1, 16:1, 14:1, 12:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1,4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:12,1:14, 1:16, 1:18, or 1:20. In certain embodiments, the ratio of the sumof compounds containing stereochemistry as depicted in 2r-(i) and2r-(ii) to the sum of compounds containing stereochemistry as depictedin 2r-(iii) and 2r-(iv) is 3:7. In certain embodiments, the ratio of thesum of compounds containing stereochemistry as depicted in 2r-(i) and2r-(ii) to the sum of compounds containing stereochemistry as depictedin 2r-(iii) and 2r-(iv) is 1:2. In certain embodiments, the ratio of thesum of compounds containing stereochemistry as depicted in 2r-(i) and2r-(ii) to the sum of compounds containing stereochemistry as depictedin 2r-(iii) and 2r-(iv) is 1:1. Exemplary such provided compounds arefound in Example 72 (Compound N-53) and Example 73 (Compound N-48).

II. Exemplary Combinations of L and R³

It will be appreciated that all combinations of the above embodiments ofL and R³ are contemplated and that the invention is not limited to thosedescribed herein. It will further be appreciated that wherein a specificL group or R³ group is described generally without specifyingstereochemistry, the present invention contemplates all embodiments ofstereochemistry associated with that group. Exemplary combinations of Land R³ embodiments are described below.

Combinations Wherein L Contains No Chiral Centers

In some embodiments, the present invention provides compounds of FormulaI and/or Formula I′, having stereochemistry as depicted in Formula 1aand/or 1b, wherein L is a C₂₋₆ hydrocarbon chain containing no chiralcenters, and wherein R³ is of Formula 1r and/or 2r.

In some embodiments, the present invention provides compounds of FormulaI and/or Formula I′, having stereochemistry as depicted in Formula 1aand/or 1b, wherein L is a C₂₋₆ hydrocarbon chain containing no chiralcenters, and wherein R³ is of Formula 1r.

In some embodiments, the present invention provides compounds of FormulaI and/or Formula I′, having stereochemistry as depicted in Formula 1aand/or 1b, wherein L is a C₂₋₆ hydrocarbon chain containing no chiralcenters, and wherein R³ is of Formula 2r.

In some embodiments, the present invention provides a compound ofFormula I or Formula I′, having stereochemistry as depicted in Formula1a, wherein L is a C₂₋₆ hydrocarbon chain containing no chiral centers,and wherein R³ is of Formula 1r.

In some embodiments, the present invention provides a compound ofFormula I or Formula I′, having stereochemistry as depicted in Formula1b, wherein L is a C₂₋₆ hydrocarbon chain containing no chiral centers,and wherein R³ is of Formula 1r.

In some embodiments, the present invention provides compounds of FormulaI or Formula I′, having stereochemistry as depicted in Formula 1a,wherein L is a C₂₋₆ hydrocarbon chain containing no chiral centers, andwherein R³ is of Formula 2r. In certain embodiments wherein L and R³ areas described above, 2r is present as a mixture of compounds containingstereochemistry as depicted in 2r-(i) and 2r-(ii). In certainembodiments wherein L and R³ are as described above, 2r is present as amixture of compounds containing stereochemistry as depicted in 2r-(iii)and 2r-(iv). In certain embodiments wherein L and R³ are as describedabove, 2r is present as a mixture of compounds containingstereochemistry as depicted in 2r-(i), 2r-(ii), 2r-(iii) and 2r-(iv). Incertain embodiments wherein L and R³ are as described above, 2r ispresent as a compound containing stereochemistry as depicted in 2r-(i),2r-(ii), 2r-(iii) or 2r-(iv). In certain embodiments, wherein L and R³are as described above, the ratio of the sum of compounds containingstereochemistry as depicted in 2r-(i) and 2r-(ii) to the sum ofcompounds containing stereochemistry as depicted in 2r-(iii) and 2r-(iv)is about 20:1, 18:1, 16:1, 14:1, 12:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1,4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:12,1:14, 1:16, 1:18, or 1:20. In certain embodiments, wherein L and R³ areas described above, the ratio of the sum of compounds containingstereochemistry as depicted in 2r-(i) and 2r-(ii) to the sum ofcompounds containing stereochemistry as depicted in 2r-(iii) and 2r-(iv)is about 3:7. In certain embodiments, wherein L and R³ are as describedabove, the ratio of the sum of compounds containing stereochemistry asdepicted in 2r-(i) and 2r-(ii) to the sum of compounds containingstereochemistry as depicted in 2r-(iii) and 2r-(iv) is 1:2. In certainembodiments, wherein L and R³ are as described above, the ratio of thesum of compounds containing stereochemistry as depicted in 2r-(i) and2r-(ii) to the sum of compounds containing stereochemistry as depictedin 2r-(iii) and 2r-(iv) is 1:1.

In some embodiments, the present invention provides a compound ofFormula I or Formula I′, having stereochemistry as depicted in Formula1b, wherein L is a C₂₋₆ hydrocarbon chain containing no chiral centers,and wherein R³ is of Formula 2r.

Combinations Wherein L Contains One Chiral Center

In some embodiments, the present invention provides compounds of FormulaI and/or Formula I′, having stereochemistry as depicted in Formula 1aand/or 1b, wherein L is a C₂₋₆ hydrocarbon chain containing a chiralcenter, and wherein R³ is of Formula 1r and/or 2r.

In some embodiments, the present invention provides a compound ofFormula I or Formula I′, having stereochemistry as depicted in Formula1a, wherein L is a C₂₋₆ hydrocarbon chain containing a chiral center,and wherein R³ is of Formula 1r. In certain embodiments, wherein R³ isas described above, L is a C₂₋₆ hydrocarbon chain containing a chiralcenter in the (R) configuration at C₁, C₂, or C₃. In certainembodiments, wherein R³ is as described above, L is a C₂₋₆ hydrocarbonchain containing a chiral center in the (S) configuration at C₁, C₂, orC₃. In certain embodiments, the invention provides compounds wherein R³is as described above and L is a C₂₋₆ hydrocarbon chain containing achiral center present in approximately a 1:1 molar ratio of (R) to (S).

In some embodiments, the present invention provides a compound ofFormula I or Formula I′, having stereochemistry as depicted in Formula1b, wherein L is a C₂₋₆ hydrocarbon chain containing a chiral center,and wherein R³ is of Formula 1r.

In some embodiments, the present invention provides a compound ofFormula I or Formula I′, having stereochemistry as depicted in Formula1a, wherein L is a C₂₋₆ hydrocarbon chain containing a chiral center,and wherein R³ is of Formula 2r.

In some embodiments, the present invention provides a compound ofFormula I or Formula I′, having stereochemistry as depicted in Formula1b, wherein L is a C₂₋₆ hydrocarbon chain containing a chiral center,and wherein R³ is of Formula 2r.

Combinations Wherein L Contains Two Chiral Centers

In some embodiments, the present invention provides compounds of FormulaI and/or Formula I′, having stereochemistry as depicted in Formula 1aand/or 1b, wherein L is a C₂₋₆ hydrocarbon chain containing two chiralcenters, and wherein R³ is of Formula 1r and/or 2r.

In some embodiments, the present invention provides a compound ofFormula I or Formula I′, having stereochemistry as depicted in Formula1a, wherein L is a C₂₋₆ hydrocarbon chain containing two chiral centers,and wherein R³ is of Formula 1r. In certain embodiments wherein R³ and Lare as described above, the two chiral centers in the C₂₋₆ hydrocarbonchain are at C₁ and C₂, wherein at least one of the two chiral centersis racemic. In certain embodiments, wherein R³ and L are as describedabove, the two chiral centers in the C₂₋₆ hydrocarbon chain are at C₁and C₃, wherein at least one of the two chiral centers is racemic. Incertain embodiments, wherein R³ and L are as described above, the twochiral centers in the C₂₋₆ hydrocarbon chain are at C₂ and C₃, whereinat least one of the two chiral centers is racemic. In certainembodiments, wherein R³ and L are as described above, the two chiralcenters in the C₂₋₆ hydrocarbon chain are at C₁ and C₂, wherein bothchiral centers are independently enantiopure. In certain embodiments,wherein R³ and L are as described above, the two chiral centers in theC₂₋₆ hydrocarbon chain are at C₁ and C₃, wherein both chiral centers areindependently enantiopure. In certain embodiments, wherein R³ and L areas described above, the two chiral centers in the C₂₋₆ hydrocarbon chainare at C₂ and C₃, wherein both chiral centers are independentlyenantiopure. In certain embodiments, wherein R³ and L are as describedabove, compounds are provided such that all possible stereoisomers of Lare present.

In some embodiments, the present invention provides a compound ofFormula I or Formula I′, having stereochemistry as depicted in Formula1b, wherein L is a C₂₋₆ hydrocarbon chain containing two chiral centers,and wherein R³ is of Formula 1r.

In some embodiments, the present invention provides a compound ofFormula I or Formula I′, having stereochemistry as depicted in Formula1a, wherein L is a C₂₋₆ hydrocarbon chain containing two chiral centers,and wherein R³ is of Formula 2r. In some embodiments, the presentinvention provides a compound of Formula I or Formula I′, havingstereochemistry as depicted in Formula 1b, wherein L is a C₂₋₆hydrocarbon chain containing two chiral centers, and wherein R³ is ofFormula 2r.

7. Regiochemistry Embodiments

In some embodiments, compounds of the present invention are provided asa mixture of one or more regioisomers (e.g., with respect to “L”). Oneof skill in the art will appreciate that all stereochemistry embodimentsdescribed herein are contemplated with respect to regioisomers and/orregioisomeric mixtures. In certain embodiments, regioisomeric mixturescontain two regioisomers present in a ratio of about 20:1, 18:1, 16:1,14:1, 12:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, or 1:1. Incertain embodiments, one or more regioisomers may be present in one ormore stereoisomeric forms, as described above.

It will apparent to one skilled in the art that regioisomeric mixturesof compounds may contain one or more regioisomers present in any ratiorelative to one another.

Exemplary such regioisomeric mixtures described herein include: (1)Example 60, wherein a composition contains a mixture of CompoundN-28:Compound N-27 in a ratio of about 7:3, wherein the chiral carbonpresent in each of Compound N-28 and Compound N-27 is present in an (R)configuration (i.e., enantiopure). In certain embodiments, the presentinvention also contemplates the chiral carbon of Compound N-28 andCompound N-27 in an (S) configuration; (2) Example 28, wherein acomposition contains a mixture of Compound N-34:Compound N-33 in a ratioof about 6:4, wherein Compound N-34 is present as an (R)(R) enantiomerand as an (S)(R) enantiomer in a ratio of about 1:1, and whereinCompound N-33 is present as an (R)(R) enantiomer and as an (S)(R)enantiomer in a ratio of about 1:1. (3) Example 58, wherein thestereochemistry is substantially similar to the stereochemistrydescribed in Example 28 above.

TABLE 1 Exemplary Compounds

A

B

C

D

E

F

G

H

I

J

K

L

M

N-1

N-2

N-3

N-4

N-5

N-6

N-7

N-8

N-9

N-10

N-11

N-12

N-13

N-14

N-15

N-16

N-17

N-18

N-19

N-20

N-21

N-22

N-23

N-24

N-25

N-26

N-27

N-28

N-29

N-30

N-31

N-32

N-33

N-34

N-35

N-36

N-37

N-38

N-39

N-40

N-41

N-42

N-43

N-44

N-45

N-46

N-47

N-48

N-49

N-50

N-51

N-52

N-53

N-54

N-55

N-56

N-57

N-58

N-59

N-60

N-61

N-62

N-63

N-64

N-65

N-66

N-67

N-68

N-69

N-70

N-71

N-72

N-73

N-74

N-75

N-76

N-77

N-78

N-79

N-80

N-81

N-82

N-83

N-84

N-85

N-86

N-87

N-88

N-89

N-90

N-91

N-92

N-93

N-94

N-95

N-96

N-97

N-98

In certain embodiments, the present invention provides any compounddepicted in Table 1, above, or a pharmaceutically acceptable saltthereof.

Unless otherwise stated, all tautomeric forms of the compounds of theinvention are within the scope of the invention. Additionally, unlessotherwise stated, structures depicted herein are also meant to includecompounds that differ only in the presence of one or more isotopicallyenriched atoms. For example, compounds having the present structuresincluding the replacement of hydrogen by deuterium or tritium, or thereplacement of a carbon by a ¹³C- or ¹⁴C-enriched carbon are within thescope of this invention. Such compounds are useful, for example, asanalytical tools, as probes in biological assays, or as therapeuticagents in accordance with the present invention. In some embodiments,the R¹ group of Formulae I and/or I′ comprises one or more deuteriumatoms. Mixtures of isomeric forms may be separated and/or purified bytechniques as would be known to one skilled in this art, including butnot limited to column chromatography.

Compounds of Formulae I, I′ and/or Ia may be provided according to thepresent invention in any of a variety of useful forms, for example aspharmaceutically acceptable salts, as particular crystal forms, etc. Insome embodiments, prodrugs of one or more compounds of the presentinvention are provided. Various forms of prodrugs are known in the art,for example as discussed in Bundgaard (ed.), Design of Prodrugs,Elsevier (1985); Widder et al. (ed.), Methods in Enzymology, vol. 4,Academic Press (1985); Kgrogsgaard-Larsen et al. (ed.); “Design andApplication of Prodrugs”, Textbook of Drug Design and Development,Chapter 5, 113-191 (1991); Bundgaard et al., Journal of Drug DeliveryReviews, 8:1-38 (1992); Bundgaard et al., J. Pharmaceutical Sciences,77:285 et seq. (1988); and Higuchi and Stella (eds.), Prodrugs as NovelDrug Delivery Systems, American Chemical Society (1975).

As described above, the present invention provides isoprenyl compoundsrelated in structure to AFC. Like AFC, in certain embodiments, isoprenylcompounds are characterized by an ability to reduce methylation of aprotein having a carboxyl-terminal

--CAAX motif, wherein C=cysteine, A=any aliphatic amino acid, and X=anyamino acid. (See Rando, U.S. Pat. No. 5,202,456). The methylationreaction which is inhibited is part of a series of post-translationalmodifications involving the --CAAX motif. These modifications includepolyisoprenylation of the cysteine of the --CAAX motif (on the sulfur),proteolysis of the carboxyl-terminal three amino acids (--AAX) andmethylation of the carboxyl group of cysteine.

In certain embodiments, provided compounds modulate a G-proteinsignaling cascade. In certain embodiments, provided compounds alter theinteractions among polyisoprenylated signal transduction proteins, suchas G-proteins and the protein regulatory targets with which theyinteract, or other intracellular signaling proteins. In certainembodiments, provided compounds modulate the inflammatory response. Incertain embodiments, provided compounds inhibit inflammation and aretherefore anti-inflammatory. In certain embodiments, provided compoundspromote inflammation and are therefore proinflammatory.

In some embodiments, provided compounds modulate levels of inflammatorymediators, such as cytokines induced by G-protein-mediated pathways(e.g., purinergic receptors). In some embodiments, provided compoundsinhibit the levels of proinflammatory mediators, such as proinflammatorycytokines. In further embodiments, provided compounds inhibit levels ofproinflammatory mediators, such as proinflammatory cytokines induced byG-protein-mediated pathways.

In some embodiments, provided compounds modulate levels of inflammatorymediators, such as cytokines induced by other signal transductionpathways [e.g., pathways involving Toll-like receptors (“TLRs”) and TNFαreceptors]. In some embodiments, provided compounds inhibit levels ofproinflammatory mediators, such as proinflammatory cytokines induced byother signal transduction pathways [e.g., pathways involving Toll-likereceptors (“TLRs”) and TNFα receptors].

In some embodiments, provided compounds inhibit levels ofproinflammatory mediators, such as proinflammatory cytokines that areinduced by chemicals such as TPA.

In some embodiments, provided compounds modulate the levels ofinflammatory mediators such as cytokines characterized using an AtopicDermatitis mouse model.

In some embodiments, provided compounds inhibit the levels ofproinflammatory mediators such as proinflammatory cytokinescharacterized using an Atopic Dermatitis mouse model.

In some embodiments, provided compounds modulate the infiltration andaccumulation of T-helper lymphocytes. In some embodiments, providedcompounds modulate T-helper lymphocytes with CD3+ marker. In someembodiments, provided compounds modulate the infiltration andaccumulation of T-helper lymphocytes characterized using a Stat3cpsoriasis mouse model. In some embodiments, provided compounds inhibitinfiltration and accumulation of T-helper lymphocytes. In someembodiments, provided compounds inhibit infiltration and accumulation ofT-helper lymphocytes with CD3+ marker. In some embodiments, providedcompounds inhibit infiltration and accumulation of T-helper lymphocytescharacterized using a Stat3c psoriasis mouse model.

In some embodiments, provided compounds inhibit methylesterificationreactions by a specific membrane associatedS-adenosylmethionine-dependent isoprenyl-S-isoprenyl methyltransferase(“ICMT”) resulting in carboxy-terminal polyisoprenoid cysteinemodifications of a number of key factors in G-protein signaling pathway.

In some embodiments, provided compounds promote inflammation and aretherefore proinflammatory.

In some embodiments, provided compounds inhibit oxidative burst fromneutrophils and are therefore anti-oxidants.

In certain embodiments, activity of provided compounds may becharacterized using a variety of in vitro or in vivo assays, involving avariety of cell-based or animal-based models. For example, data fromexemplary assays for: Edema, Erythema and/or Inhibition ofMyeloperoxidase; Inflammatory Cytokines; Stat3c-Psoriasis Mouse Model;Inhibition of Methylesterification Reactions; and Inhibition ofOxidative Burst are each described below.

Edema, Erythema and/or Inhibition of Myeloperoxidase (MPO)

Ability of provided compounds to modulate inflammatory responses may beassessed, for example, using assays that assess edema, erythema, and/orinhibition of myeloperoxidase (“MPO”) as described, for example, inExample 79.

In certain embodiments, provided compounds are considered to beinhibitors of inflammation when they show a percent inhibition in anedema assay of at least about 30, 35, 40, 50, 60, 70, 80, 90 or 95%, forexample when provided at a dose 0.8 mg/20 μL. In certain embodiments,provided compounds are considered to be inhibitors of inflammation whenthey show a percent inhibition in an edema assay of at least about 5,10, 15, 20, 25, 30, 35, 40, 50, 60, 70, or 80%, for example whenprovided at a dose of 0.2 mg/20 μL. In certain embodiments, providedcompounds are considered to be inhibitors of inflammation when theyresult in an ED₅₀ in an edema assay of at least about 0.5, 1, 1.5, 2,2.5, 3, 3.5, 4, 4.5-fold lower than that observed with AFC. In certainembodiments, provided compounds are considered to be proinflammatorywhen they show percent inhibition in an edema assay of at least about(−)10, (−)20, (−)30, (−)40, (−)50, (−)55, (−)60, (−)65, (−)70, (−)75,(−)80, (−)85, (−)90 or (−)95%, for example when provided at a dose of0.8 mg/20 μl.

In certain embodiments, provided compounds are considered to beinhibitors of inflammation when they show a percent inhibition in anerythema assay of at least about 25, 30, 35, 40, 50, 60, 70, 80, 90 or95%, for example when provided at a dose of 0.8 mg/20 μL. In certainembodiments, provided compounds are considered to be inhibitors ofinflammation when they show a percent inhibition in an erythema assay ofat least about 5, 10, 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90 or 95%,for example when provided at a dose of 0.2 mg/20 μL. In certainembodiments, provided compounds are considered to be inhibitors ofinflammation when they result in an ED₅₀ in an erythema assay of atleast about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5-fold lower than thatobserved with AFC. In certain embodiments, provided compounds areconsidered to be proinflammatory when they show percent inhibition in anerythema assay of at least about (−)10, (−)20, (−)30, (−)40, (−)50,(−)55, (−)60, (−)65, (−)70, (−)75, (−)80, (−)85, (−)90 or (−)95%, forexample when provided at a dose of 0.8 mg/20 μl.

In certain embodiments, provided compounds are considered to beinhibitors of inflammation when they show a percent inhibition in an MPOactivity assay of at least about 60, 70, 80, 90 or 95%, for example whenprovided at a dose of 0.8 mg/20 μL. In certain embodiments, providedcompounds are considered to be inhibitors of inflammation when they showa percent inhibition in an MPO activity assay of at least about 5, 10,15, 20, 25, 30, 35, 40, 50, 60, 70, or 80%, for example when provided ata dose of 0.2 mg/20 μL. In certain embodiments, provided compounds areconsidered to be inhibitors of inflammation when they result in an ED₅₀in an MPO activity assay of at least about 0.5, 1, 1.5, 2, 2.5, 3, 3.5,4, 4.5, 5, 5.5, 6 or 6.5-fold lower than that observed with AFC. Incertain embodiments, provided compounds are considered to beproinflammatory when they show percent inhibition in an MPO activityassay of at least about (−)10, (−)20, (−)30, (−)40, (−)50, (−)55, (−)60,(−)65, (−)70, (−)75, (−)80, (−)85, (−)90 or (−)95%, for example whenprovided at a dose of 0.8 mg/20 μl.

Inflammatory Cytokines

Ability of provided compounds to modulate inflammatory responses may beassessed for example, using assays that measure the levels ofinflammatory cytokines, for example, TNF-α, IL-1β, IL-8/KC, or IL-6,that can be determined using inflammatory models [e.g., TPA-inducedmouse ear inflammatory model as described in Example 80;LPS-TLR4-induced cytokine release inflammatory model in HumanMicrovascular Endothelial cell lines (“HMEC-1”) as described in Example81; ATPγS-purinergic receptor-induced cytokine release inflammatorymodel in Human Microvascular Endothelial cell lines (“HMEC-1”) asdescribed in Example 82; TPA-induced cytokine release inflammatory modelin Normal Human Epidermal Keratinocyte cell lines (“NHEK”) as describedin Example 83; TNFα-induced cytokine release inflammatory model in HumanUmbilical Vein Endothelial cell lines (“HUVEC”) as described in Example84; or an Ovalbumin-induced flaky tail Atopic Dermatis mouse model asdescribed in Example 85].

(i) TPA-Induced Mouse Ear Inflammatory Model

In certain embodiments, provided compounds are considered inhibitors ofinflammation when they show a percent inhibition of cytokine release ina TPA-induced mouse ear inflammatory model of at least about 5, 10, 15,20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 95%, forexample when provided at a dosage of 0.25%. In certain embodiments,provided compounds are considered inhibitors of inflammation when theyresult in an ED₅₀ in a TPA-induced mouse ear inflammatory model of atleast about 0.01, 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35 or 0.40 μgcytokine/mouse ear, for example when provided at a dosage of 0.25%. Incertain embodiments, provided compounds are considered inhibitors ofinflammation when they show a percent inhibition of cytokine release ina TPA-induced mouse ear inflammatory model of at least about 5, 10, 15,20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 95%, forexample when provided at a dosage of 0.50%. In certain embodiments,provided compounds are considered inhibitors of inflammation when theyresult in an ED₅₀ in a TPA-induced mouse ear inflammatory model of atleast about 0.01, 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35 or 0.40 μgcytokine/mouse ear, for example when provided at a dosage of 0.50%. Incertain embodiments, provided compounds are considered inhibitors ofinflammation when they show a percent inhibition of cytokine release ina TPA-induced mouse ear inflammatory model of at least about 5, 10, 15,20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 95%, forexample when provided at a dosage of 1.00%. In certain embodiments,provided compounds are considered inhibitors of inflammation when theyresult in an ED₅₀ in a TPA-induced mouse ear inflammatory model of atleast about 0.01, 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35 or 0.40 μgcytokine/mouse ear, for example, when provided at a dosage of 1.00%.

(ii) LPS-TLR4-Induced Cytokine Release Inflammatory Model

In certain embodiments, provided compounds are considered inhibitors ofinflammation when they show a percent inhibition of cytokine release ina LPS-TLR4-induced cytokine release model, as determined using HMEC-1cells of at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60,65, 70, 75, 80, 85, 90 or 95%, for example when provided at a dosage of0.25%. In certain embodiments, provided compounds are consideredinhibitors of inflammation when they result in an ED₅₀ in aLPS-TLR4-induced cytokine release model, as determined using HMEC-1cells, of at least about 0.01, 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35or 0.40 μg cytokine/mouse ear, for example when provided at a dosage of0.25%. In certain embodiments, provided compounds are consideredinhibitors of inflammation when they show a percent inhibition ofcytokine release in a LPS-TLR4-induced cytokine release model, asdetermined using HMEC-1 cells, of at least about 5, 10, 15, 20, 25, 30,35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 95%, for example whenprovided at a dosage of 0.50%. In certain embodiments, providedcompounds are considered inhibitors of inflammation when they result inan ED₅₀ in a LPS-TLR4-induced cytokine release model, as determinedusing HMEC-1 cells, of at least about 0.01, 0.05, 0.10, 0.15, 0.20,0.25, 0.30, 0.35 or 0.40 μg cytokine/mouse ear, for example whenprovided at a dosage of 0.50%. In certain embodiments, providedcompounds are considered inhibitors of inflammation when they show apercent inhibition of cytokine release in a LPS-TLR4-induced cytokinerelease model, as determined using HMEC-1 cells, of at least about 5,10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or95%, for example when provided at a dosage of 1.00%. In certainembodiments, provided compounds are considered inhibitors ofinflammation when they result in an ED₅₀ in a LPS-TLR4-induced cytokinerelease model, as determined using HMEC-1 cells, of at least about 0.01,0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35 or 0.40 μg cytokine/mouse ear,for example, when provided at a dosage of 1.00%.

(iii) ATPγS-Purinergic Receptor-Induced Cytokine Release InflammatoryModel

In certain embodiments, provided compounds are considered inhibitors ofinflammation when they show a percent inhibition of cytokine release inan ATPγS-purinergic receptor-induced cytokine release model, asdetermined using HMEC-1 cells of at least about 5, 10, 15, 20, 25, 30,35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 95%, for example whenprovided at a dosage of 0.25%. In certain embodiments, providedcompounds are considered inhibitors of inflammation when they result inan ED₅₀ in an ATPγS-purinergic receptor-induced cytokine release model,as determined using HMEC-1 cells, of at least about 0.01, 0.05, 0.10,0.15, 0.20, 0.25, 0.30, 0.35 or 0.40 μg cytokine/mouse ear, for examplewhen provided at a dosage of 0.25%. In certain embodiments, providedcompounds are considered inhibitors of inflammation when they show apercent inhibition of cytokine release in an ATPγS-purinergicreceptor-induced cytokine release model, as determined using HMEC-1cells, of at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60,65, 70, 75, 80, 85, 90 or 95%, for example when provided at a dosage of0.50%. In certain embodiments, provided compounds are consideredinhibitors of inflammation when they result in an ED₅₀ in anATPγS-purinergic receptor-induced cytokine release model, as determinedusing HMEC-1 cells, of at least about 0.01, 0.05, 0.10, 0.15, 0.20,0.25, 0.30, 0.35 or 0.40 μg cytokine/mouse ear, for example whenprovided at a dosage of 0.50%. In certain embodiments, providedcompounds are considered inhibitors of inflammation when they show apercent inhibition of cytokine release in an ATPγS-purinergicreceptor-induced cytokine release model, as determined using HMEC-1cells, of at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60,65, 70, 75, 80, 85, 90 or 95%, for example when provided at a dosage of1.00%. In certain embodiments, provided compounds are consideredinhibitors of inflammation when they result in an ED₅₀ in anATPγS-purinergic receptor-induced cytokine release model, as determinedusing HMEC-1 cells, of at least about 0.01, 0.05, 0.10, 0.15, 0.20,0.25, 0.30, 0.35 or 0.40 μg cytokine/mouse ear, for example, whenprovided at a dosage of 1.00%.

(iv) TPA-Induced Cytokine Release Inflammatory Model

In certain embodiments, provided compounds are considered inhibitors ofinflammation when they show a percent inhibition of cytokine release ina TPA-induced cytokine release model, as determined using NHEK cells ofat least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70,75, 80, 85, 90 or 95%, for example when provided at a dosage of 0.25%.In certain embodiments, provided compounds are considered inhibitors ofinflammation when they result in an ED₅₀ in a TPA-induced cytokinerelease model, as determined using NHEK cells, of at least about 0.01,0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35 or 0.40 μg cytokine/mouse ear,for example when provided at a dosage of 0.25%. In certain embodiments,provided compounds are considered inhibitors of inflammation when theyshow a percent inhibition of cytokine release in a TPA-induced cytokinerelease model, as determined using NHEK cells, of at least about 5, 10,15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 95%,for example when provided at a dosage of 0.50%. In certain embodiments,provided compounds are considered inhibitors of inflammation when theyresult in an ED₅₀ in a TPA-induced cytokine release model, as determinedusing NHEK cells, of at least about 0.01, 0.05, 0.10, 0.15, 0.20, 0.25,0.30, 0.35 or 0.40 μg cytokine/mouse ear, for example when provided at adosage of 0.50%. In certain embodiments, provided compounds areconsidered inhibitors of inflammation when they show a percentinhibition of cytokine release in a TPA-induced cytokine release model,as determined using NHEK cells, of at least about 5, 10, 15, 20, 25, 30,35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 95%, for example whenprovided at a dosage of 1.00%. In certain embodiments, providedcompounds are considered inhibitors of inflammation when they result inan ED₅₀ in a TPA-induced cytokine release model, as determined usingNHEK cells, of at least about 0.01, 0.05, 0.10, 0.15, 0.20, 0.25, 0.30,0.35 or 0.40 μg cytokine/mouse ear, for example, when provided at adosage of 1.00%.

(v) TNFα-Induced Cytokine Release Inflammatory Model

In certain embodiments, provided compounds are considered inhibitors ofinflammation when they show a percent inhibition of cytokine release ina TNFα-induced cytokine release model, as determined using HUVEC cellsof at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70,75, 80, 85, 90 or 95%, for example when provided at a dosage of 0.25%.In certain embodiments, provided compounds are considered inhibitors ofinflammation when they result in an ED₅₀ in a TNFα-induced cytokinerelease model, as determined using HUVEC cells, of at least about 0.01,0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35 or 0.40 μg cytokine/mouse ear,for example when provided at a dosage of 0.25%. In certain embodiments,provided compounds are considered inhibitors of inflammation when theyshow a percent inhibition of cytokine release in a TNFα-induced cytokinerelease model, as determined using HUVEC cells, of at least about 5, 10,15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 95%,for example when provided at a dosage of 0.50%. In certain embodiments,provided compounds are considered inhibitors of inflammation when theyresult in an ED₅₀ in a TNFα-induced cytokine release model, asdetermined using HUVEC cells, of at least about 0.01, 0.05, 0.10, 0.15,0.20, 0.25, 0.30, 0.35 or 0.40 μg cytokine/mouse ear, for example whenprovided at a dosage of 0.50%. In certain embodiments, providedcompounds are considered inhibitors of inflammation when they show apercent inhibition of cytokine release in a TNFα-induced cytokinerelease model, as determined using HUVEC cells, of at least about 5, 10,15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 95%,for example when provided at a dosage of 1.00%. In certain embodiments,provided compounds are considered inhibitors of inflammation when theyresult in an ED₅₀ in a TNFα-induced cytokine release model, asdetermined using HUVEC cells, of at least about 0.01, 0.05, 0.10, 0.15,0.20, 0.25, 0.30, 0.35 or 0.40 μg cytokine/mouse ear, for example whenprovided at a dosage of 1.00%.

(vi) Ovalbumin-Induced Flaky Tail Atopic Dermatis Mouse Model

In certain embodiments, provided compounds are considered inhibitors ofinflammation when they show a percent inhibition of cytokine release inan Ovalbumin-induced Atopic Dermatitis mouse model, of at least about 5,10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or95%, for example when provided at a dosage of 0.25%. In certainembodiments, provided compounds are considered inhibitors ofinflammation when they result in an ED₅₀ in an Ovalbumin-induced AtopicDermatitis mouse model, of at least about 0.01, 0.05, 0.10, 0.15, 0.20,0.25, 0.30, 0.35 or 0.40 μg cytokine/mouse ear, for example whenprovided at a dosage of 0.25%. In certain embodiments, providedcompounds are considered inhibitors of inflammation when they show apercent inhibition of cytokine release in an Ovalbumin-induced AtopicDermatitis mouse model, of at least about 5, 10, 15, 20, 25, 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 95%, for example when providedat a dosage of 0.50%. In certain embodiments, provided compounds areconsidered inhibitors of inflammation when they result in an ED₅₀ in aan Ovalbumin-induced Atopic Dermatitis mouse model, of at least about0.01, 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35 or 0.40 μg cytokine/mouseear, for example when provided at a dosage of 0.50%. In certainembodiments, provided compounds are considered inhibitors ofinflammation when they show a percent inhibition of cytokine release inan Ovalbumin-induced Atopic Dermatitis mouse model, of at least about 5,10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or95%, for example when provided at a dosage of 1.00%. In certainembodiments, provided compounds are considered inhibitors ofinflammation when they result in an ED₅₀ in an Ovalbumin-induced AtopicDermatitis mouse model, of at least about 0.01, 0.05, 0.10, 0.15, 0.20,0.25, 0.30, 0.35 or 0.40 μg cytokine/mouse ear, for example, whenprovided at a dosage of 1.00%.

Stat3c-Psoriasis Mouse Model

For example, ability of provided compounds to modulate inflammatoryresponses may be assessed for example, using assays that measure thelevels of CD3+ T-helper cells, that can be determined using mousemodels, for example, a Stat3c-psoriasis mouse model, as described inExample 86. In certain embodiments, provided compounds are consideredinhibitors of the infiltration and accumulation of CD3+ T-helper cellswhen they show a percent reduction of the number of T-helper cells of atleast about 20, 25, 30, 35, 40, 50, 60, 70, 75, 80, 85, 90, 95 or 100%,for example when provided at a concentration of at least 0.3%.

Inhibition of Methylesterification Reactions

For example, ability of provided compounds to inhibitmethylesterification reactions by ICMT may be assessed, for example,using assays that measure the reduction of methylated acetyl farnesylcysteine, an ICMT substrate as described for example in Example 87. Incertain embodiments, provided compounds are considered inhibitors ofICMT when they show a percent reduction of methylatedacetyl-farnesyl-cysteine, as ICMT substrate of at least about 30, 35,40, 50, 60, 70, 75, 80, 85, 90, 95 or 100%, for example when provided ata concentration of 25 μM.

Inhibition of Oxidative Burst

For example, ability of provided compounds to inhibit oxidative burstfrom neutrophils may be assessed, for example, using assays that measurethe reduction of superoxide formation, as described for example inExample 88. In certain embodiments, provided compounds are consideredinhibitors of oxidative burst from neutrophils when they show a percentreduction of superoxide formation of at least about 30, 35, 40, 50, 60,70, 75, 80, 85, 90, 95 or 100%, for example when provided at aconcentration of 25 μM.

2. Methods of Syntheses

The present invention provides methods of preparing compounds providedherein. As will be appreciated by one of skill in the art, the syntheticmethods described herein may be modified without departing from thescope of the present invention. For example, different startingmaterials and/or different reagents may be used in the inventivesynthetic methods.

The present invention provides a process for preparing an N-substitutedfarnesyl cysteine analog with a terminal carboxylic acid. In certainembodiments, the inventive compounds are prepared as shown in the schemebelow.

To begin, a suitable compound 1 is reacted with a suitable electrophiliccompound 2. In certain embodiments, the suitable compound 1 is S-trans,trans-farnesyl-L-cysteine. In certain embodiments, the electrophiliccompound 2 is an anhydride. Exemplary anhydrides include succinicanhydride, maleic anhydride, 3-methylenedihydro-2,4-furandione, glutaricanhydride, N-phthaloyl-glutamic anhydride. In certain embodiments, theelectrophilic compound 2 is an isocyanate. In certain embodiments, theisocyanate is ethyl-3-isocyanato-propionate. In certain embodiments, theelectrophilic compound 2 is an activated ester of an acid. Exemplaryactivated esters of an acid include maleamic acid, mono-ethyl fumarateand BOC-glutamine. In certain embodiments, the electrophilic compound 2is an acid chloride. Exemplary acid chlorides include adipoyl chloride,maleyl chloride, and sebacoyl chloride, etc. In certain embodiments, theelectrophilic compound 2 is a sulfonyl chloride. Exemplary sulfonylchlorides include cyclopropane sulfonyl chloride, ethyl3-(chlorosulfonyl)propanoate, and ethyl 2-(chlorosulfonyl)benzoate, etc.In certain embodiments, the electrophilic compound 2 is an activatedacid. Exemplary activated acids include an acid that has been treatedwith an activating agent. One skilled in the art will be able toidentify an appropriate activating agent from exemplary activatingagents, including but not limited to the list as defined herein. Thereaction is typically performed in the presence of a suitable base toform a compound of formula I. In certain embodiments, the base is K₂CO₃.The reaction is typically performed in a suitable solvent. In certainembodiments, the suitable solvent is a mixture of polar, aproticsolvents. In certain embodiments, whether used alone or as part of amixture, the polar, aprotic solvents include DMF, DCM, NMP and THF.

In the above-described scheme and/or steps, the R¹, R², and R³, groupsof the various formulae are as described herein.

In some embodiments, the present invention provides processes forpreparing exemplary compounds of the present invention (e.g., CompoundN-24, Compound N-38, and Compound N-34). In some embodiments, certaincompounds of the present invention are prepared according to the schemebelow.

To begin, a suitable compound 3 is reacted with a suitable electrophiliccompound 4 to give compound 5. In certain embodiments, the suitablecompound 3 is S-trans, trans-farnesyl-L-cysteine methyl ester. Reactionof compound 5 with hydrazine results in compound 6. In certainembodiments, a suitable solvent includes a mixture of polar, aproticsolvents. In certain embodiments, whether used alone or as part of amixture, the polar, aprotic solvents include, but are not limited to,DMF, DCM, NMP and THF.

In some embodiments, the present invention provides processes forpreparing exemplary compounds of the present invention (e.g., CompoundN-67, Compound N-68, N-69 and Compound N-70). In some embodiments,certain compounds of the present invention are prepared according to thescheme below.

Commercially available 2-chlorotritylchloride resin is coupled toFmoc-Cys(SStBu)-OH. Reductive removal of the dithio-tert-butylprotecting group from 7 with dithiothreitol is followed by coupling ofthe desired R³ side chain to the free thiol, using alkyl halide toafford intermediate 8. The Fmoc protecting group is typically removedwith 20% piperidine/DMF; followed by coupling an appropriateFmoc-protected acid with the resulting free amine is accomplished usingan activating agent. One skilled in the art will be able to identify anappropriate activating agent, which includes, but is not limited to, thelist as defined herein. The Fmoc protecting group is removed with 20%piperidine/DMF. Coupling of the selected N-protecting reagents with theresulting free amine is typically performed in the presence of asuitable base to form a compound of formula 9. In certain embodiments,the base is K₂CO₃. The polymer-bound prenylcysteine analog 10 istypically released from the resin using optimized cleavage conditions[e.g., stir 3×1 minute in 1% TFA CH₂Cl₂ solution].

In some embodiments, the present invention provides processes forpreparing exemplary compounds of the present invention (e.g., CompoundN-54, Compound N-32, N-78 and Compound N-77). In some embodiments,certain compounds of the present invention are prepared according to thescheme below.

To begin, a suitable compound 3 is reacted with a suitable Fmocprotected amino acid to give a coupled product 11. In certainembodiments, the suitable compound 3 is S-trans,trans-farnesyl-L-cysteine methyl ester. In certain embodiments, thesuitable amino acid is an α-amino acid (n=0). In certain embodiments,the suitable amino acid is a β-amino acid (n=1). In certain embodiments,the suitable amino acid is a γ-amino acid (n=2). In certain embodiments,the suitable amino acid is a δ-amino acid (n=3). Deprotection ofcompound 11 followed by reaction with a suitable electrophilic to affordcompound 12. In certain embodiments, the electrophilic compound is ananhydride. In certain embodiments, the electrophilic compound is anisocyanate. In certain embodiments, the isocyanate is ethylisocyanate.In certain embodiments, the electrophilic compound is an anhydride, acidchloride or activated acid. Exemplary anhydrides include aceticanhydride.

3. Compositions and Formulations

The present invention provides compositions comprising isoprenylcompounds as described herein. In some embodiments, providedcompositions contain additional components. In some embodiments, allsuch additional components are pharmaceutically acceptable and providedcompositions are pharmaceutical compositions. In some embodiments, allsuch additional components are cosmetically acceptable and providedcompositions are cosmetic compositions. In some embodiments, all suchadditional components are cosmeceutically acceptable and providedcompositions are cosmeceutical compositions.

In some embodiments, pharmaceutical, cosmetic or cosmeceuticalcompositions of the present invention comprise an isoprenyl compound, apharmaceutically acceptable inert ingredient (e.g., a carrier) andoptionally an additional active ingredient. In certain embodiments, theisoprenyl compound is a compound of Formulae I, I′ and/or Ia. In certainembodiments, the isoprenyl compound is a compound of Formulae I, I′and/or Ia. In certain embodiments, the isoprenyl compound is a compoundof Formulae I, I′ and/or Ia.

In general, one or more compounds of the present invention may beformulated into pharmaceutical compositions that include at least oneprovided compound of the present invention together with one or morepharmaceutically acceptable carriers, including excipients, such asdiluents, binders and the like, and additives, such as stabilizingagents, preservatives, solubilizing agents, and buffers, as desired.Formulation excipients may include polyvinylpyrrolidone, gelatin,hydroxy cellulose, acacia, polyethylene glycol, manniton, sodiumchloride and sodium citrate. In some embodiments, inventive compositionscontain a pharmaceutically acceptable carrier. In some embodiments, thecompositions of the present invention include a cosmetically acceptablecarrier. In some embodiments, the compositions of the present inventioninclude a cosmeceutically acceptable carrier.

Pharmaceutical carriers are typically of sufficiently high purity andsufficiently low toxicity to render it suitable for administration tothe subject being treated. Pharmaceutical carriers further maintainstability and bioavailability of an active agent (e.g., a isoprenylcompound of the present invention). Pharmaceutical carriers can beliquid or solid and are selected with the planned manner ofadministration in mind, to provide for the desired bulk, consistency,etc., when combined with an active agent and other components of a givencomposition.

A carrier in certain compositions according to the present invention mayinclude liquid and, in particular may comprise a buffered, isotonic,aqueous solution.

A carrier, including a pharmaceutically acceptable carrier, may be, orinclude, an excipient, such as a diluent, binder (e.g., binding agent)and the like, and or an additive, such as a stabilizing agent,preservative, solubilizing agent, and/or buffer as hereafter described.Pharmaceutical carriers include, without limitation, a binding agent(e.g., hydroxypropyl methylcellulose, polyvinylpyrrolidone, orpregelatinised maize starch, etc.); a filler (e.g., calcium hydrogenphosphate calcium sulfate, ethyl cellulose, gelatin, lactose and othersugars, microcrystalline cellulose, pectin, polyacrylates, etc.); adisintegrant (e.g., glycolate, sodium starch, starch, etc.); a lubricant(e.g., colloidal silicon dioxide, corn starch, hydrogenated vegetableoils, polyethylene glycols, magnesium stearate, metallic stearates,silica, sodium benzoate, sodium acetate, stearic acid, talc, etc.); or awetting agent (e.g., sodium lauryl sulphate, etc.). Additionalpharmaceutically acceptable carriers include, for example, petroleumjelly (Vaseline™), and petroleum.

Additional suitable carriers for the compositions of the presentinvention include, but are not limited to, alcohols, amyloses, animaloil, anti-irritants, chelating agents, colorants, deodorant agents,emulsifiers, fragrances, gelatins, hair conditioning agents,hydroxymethylcelluloses, magnesium stearates moisturizing agents (e.g.,humectants), microcrystalline, mineral oil, natural polymers (e.g.,collagen, gum arabic, polyols, and xanthanes, and the like), organic,ozocerite wax, and inorganic waxes, paraffin, penetration enhancers, pHadjusting agents, preservatives, propellants, salt solutions, silicicacids, surfactants talcs, solubilizing agents, thickeners, viscousparaffins, and water, and combinations thereof. In some embodiments,isoprenyl compounds of the present invention act as acceptablecarrier(s) and/or excipient(s). In certain embodiments, AFC acts as anacceptable carrier and/or excipient. In some embodiments, it may bedesirable to use the carriers in cosmetic compositions, as described inthe CTFA International Cosmetic Ingredient Dictionary and Handbook, 8thedition, edited by Wenninger and Canterbery, (The Cosmetic, Toiletry,and Fragrance Association, Inc., Washington, D.C., 2000), which isherein incorporated by reference. Also included are the carriersdescribed hereinabove.

In some embodiments, pharmaceutically acceptable carriers of thecomposition include a sustained release or delayed release carrier. Suchcarriers can be any material capable of sustained or delayed release ofisoprenyl compounds to provide a more efficient administration resultingin less frequent and/or decreased dosage of isoprenyl compounds, ease ofhandling, and extended or delayed effects on diseases, disorders,conditions, syndromes, and the like, being treated, prevented orpromoted. Non-limiting examples of such carriers include liposomes,microsponges, microspheres, or microcapsules of natural and syntheticpolymers and the like. Liposomes which may enhance the localizeddelivery of the compounds of the inventive composition within skinlayers, may be formed from a variety of phospholipids, such ascholesterol, stearylamines or phosphatidylcholines.

For injection or other liquid administration formulations, watercontaining at least one or more buffering constituents is commonlyutilized, and stabilizing agents, preservatives and solubilizing agentsmay also be employed. In some embodiments, a provided pharmaceuticalcomposition is or comprises an isotonic solution.

For solid administration formulations, any of a variety of thickening,filler, bulking and carrier additives may be employed, such as starches,sugars, fatty acids and the like. Topical compositions of the presentinvention can be applied locally to the skin or mucosa and may be in anyform including solutions, oils, creams, ointments, gels, lotions,shampoos, milks, cleansers, moisturizers, sprays, skin patches and thelike.

For most pharmaceutical formulations, non-active ingredients willconstitute the greater part, by weight or volume, of the preparation.For pharmaceutical formulations, it is also contemplated that any of avariety of measured-release, slow-release or time-release formulationsand additives may be employed, so that the dosage may be formulated soas to effect delivery of a provided compound over a period of time. Forexample, gelatin, sodium carboxymethylcellulose and/or other cellulosicexcipients may be included to provide time-release or slower-releaseformulations, especially for administration by subcutaneous andintramuscular injection.

In practical use, inventive compounds can be combined as the activeingredient in an admixture with a pharmaceutical carrier according toconventional pharmaceutical compounding techniques. Pharmaceuticalcompositions for the present invention may be formulated for delivery byany of a variety of routes including, for example, oral, parenteral(including intravenous), urethral, vaginal, nasal, topical (e.g.,dermal, transdermal), pulmonary, deep lung, inhalation, buccal,sublingual routes, or the like.

In preparing compositions compositions containing isoprenyl compoundsfor cutaneous administration, such as topical (i.e., local), suchcompositions can include pharmaceutical carriers (e.g., sterile andnon-sterile aqueous solutions, non-aqueous solutions in common solventssuch as alcohols, or solutions of isoprenyl compounds in liquid or solidoil bases). Such pharmaceutical carrier solutions also can containbuffers, diluents and other suitable additives.

In preparing compositions containing isoprenyl compounds for parenteraladministration (e.g., intramuscular or subcutaneous administration),such compositions can include pharmaceutical carriers (e.g., sterile andnon-sterile aqueous solutions, non-aqueous solutions in common solventssuch as alcohols, or solutions of isoprenyl compounds in liquid or solidoil bases). Such pharmaceutical carrier solutions also can containbuffers, diluents and other suitable additives.

Representative compositions suitable for oral use include, for example,mouthwash, rinse, oral spray, suspension, dental gel, and the like.Typical oral carriers known in the art may be used in the presentinvention. The preferred pharmaceutical and/or cosmetic carriers arewater, ethanol, and water-ethanol mixtures. The water-ethanol mixturesare generally employed in a weight ratio from about 1:1 to about 20:1,preferably from about 3:1 to about 20:1, and most preferably from about3:1 to about 10:1, respectively. The pH value of the oral vehicle isgenerally from about 4 to about 7, and preferably from about 5 to about6.5. An oral topical vehicle having a pH value below about 4 isgenerally irritating to the oral cavity and an oral vehicle having a pHvalue greater than about 7 generally results in an unpleasant mouthfeel.

Oral topical inventive compositions may also contain conventionaladditives normally employed in those products. Conventional additives asdescribed herein include a coloring agents, emulsifiers, fluorineproviding compounds, humectants, sweetening agents, and pH adjustingagents, provided that such additives do not interfere with thetherapeutic, cosmetically, or cosmeceutically beneficial properties ofinventive compositions. Additional ingredients that may be used incompositions of the present invention include fluorine providingcompounds, additional active ingredients, new excipients, protectives,and demulcents, as described herein.

Fluorine providing compounds may be fully or slightly water soluble andare characterized by their ability to release fluoride ions or fluoridecontaining ions in water and by their lack of reaction with othercomponents in the composition. Typical fluorine providing compoundsinclude alkali metal fluorides, inorganic fluoride salts such aswater-soluble alkali metal, alkaline earth metal, heavy metal salts, forexample, aluminum mono- and di-fluorophosphates, ammonium fluoride,ammonium fluorosilicate, barium fluoride, cuprous fluoride, fluorinatedsodium calcium pyrophosphate, potassium fluoride, sodium fluoride,sodium fluorosilicate, sodium fluorozirconate, sodiummonofluorophosphate, stannic fluoride, stannous fluoride and zincfluoride, monofluorophosphates, such as sodium and stannous fluoride,sodium monofluorophosphate, tin fluoride and combinations thereof.

Amounts of fluorine providing compounds present in oral, topicalinventive compositions provided herein depend upon the type of fluorineproviding compound employed, solubility of the fluorine compound, andthe nature of the final oral inventive composition. Amount of fluorineproviding compounds used must be a nontoxic amount. In general, fluorineproviding compounds when used will be present in an amount up to about1%, from about 0.001% to about 0.1%, and from about 0.001% to about0.05%, by weight of oral topical inventive compositions provided herein.

Typical sweetening agents (sweeteners) that are well known in the artinclude those that are both natural and artificial sweeteners, may beemployed. Sweetening agent used may be selected from a wide range ofmaterials including water-soluble sweetening agents, water-solubleartificial sweetening agents, water-soluble sweetening agents derivedfrom naturally occurring water-soluble sweetening agents, dipeptidebased sweetening agents, and protein based sweetening agents, includingmixtures thereof.

In some embodiments, compositions of the present invention can furtherinclude one or more additional (“compatible”, as defined herein) activeingredients which are aimed at providing compositions with anotherpharmaceutical, cosmetic, or cosmeceutical effect, in addition to thatprovided by an isoprenyl compound of inventive compositions providedherein.

Additional active ingredients according to the present inventioninclude, without limitation, one or more, in any combination, of aprotective agent, an emollient, an astringent, an irritant, akeratolytic, a sun screening agent, a sun tanning agent, an antibioticagent, an antifungal agent, an antiviral agent, an antiprotozoal agent,an anesthetic agent, a steroidal anti-inflammatory agent, anon-steroidal anti-inflammatory agent, an antipruritic agent, ananti-oxidant agent, a chemotherapeutic agent, an anti-histamine agent, avitamin, a hormone, an anti-dandruff agent, an anti-wrinkle agent, ananti-skin atrophy agent, a sclerosing agent, a cleansing agent, acaustic agent and a hypo-pigmenting agent.

In some embodiments, at least one isoprenyl compound of compositionsprovided herein is an active ingredient.

Compositions according to the present invention, which further includeone or more additional active ingredients, can therefore be furtherefficiently used, in addition to their use as a treatment for anepithelial-related condition, in the treatment of any medical, cosmeticand/or cosmeceutical condition in which applying the additional activeingredient is beneficial.

Protectives as described herein may take the form of dusting powders,adsorbents, mechanical protective agents, and plasters. Dusting powdersare relatively inert and insoluble materials that are used to cover andprotect epithelial surfaces, ulcers and wounds. Usually, thesesubstances are finely subdivided powders that absorb moisture and canact as a dessicant. The absorption of skin moisture decreases frictionand also discourages certain bacterial growth. Some of the materialsused as protective adsorbents include bentonite, insoluble salts ofbismuth, boric acid, calcium carbonate, (precipitated), cellulose, cornstarch, magnesium stearate, talc, titanium dioxide, zinc oxide, and zincstearate.

In some embodiments, protectives also can be administered to the skin toform an adherent, continuous film that may be flexible or semi-rigiddepending on the materials and the formulations as well as the manner inwhich they are applied. This material may serve several purposesincluding providing occlusion from the external environment, providingchemical support, and serving as vehicles for other medicaments.

In some embodiments, protectives included in compositions of the presentinvention are demulcents. Demulcents often are applied to the surface ina viscid, sticky preparation that covers the area readily and may bemedicated. A number of chemical substances possess demulcent properties.

In practical use, provided compounds herein can be combined as an activeingredient in an admixture with a pharmaceutical carrier according toconventional pharmaceutical compounding techniques. The carrier may takea wide variety of forms depending on the form of preparation desired foradministration, for example, oral, parenteral (including intravenous),urethral, vaginal, nasal, dermal, transdermal, pulmonary, deep lung,inhalation, buccal, sublingual, or the like.

In preparing the compositions for oral dosage form, any of the usualpharmaceutical media may be employed, such as, for example, water,glycols, oils, alcohols, flavoring agents, preservatives, coloringagents and the like in the case of oral liquid preparations, such as,for example, suspensions, elixirs and solutions; or carriers such asstarches, sugars, microcrystalline cellulose, diluents, granulatingagents, lubricants, binders, disintegrating agents and the like in thecase of oral solid preparations such as, for example, powders, hard andsoft capsules and tablets. Tablets, pills, capsules, and the like mayalso contain a binder such as gum tragacanth, acacia, corn starch orgelatin; excipients such as dicalcium phosphate; a disintegrating agentsuch as corn starch, potato starch or alginic acid; a lubricant such asmagnesium stearate; and/or a sweetening agent such as sucrose, lactoseor saccharin. Capsules may contain, in addition to materials of theabove type, a liquid carrier such as a fatty oil.

In some embodiments, an isoprenyl compound, carrier and, optionally,additional active ingredients are formed into a composition in the formof a solution, emulsion or gel suspension, as will be further describedherein.

In some embodiments, an isoprenyl compound, a pharmaceutical or cosmeticcarrier and, optionally, one or more additional active ingredients, arein the form of a solution. A solution can be prepared by mixing a soluteor dissolved substance (such as a isoprenyl compound of the inventionand, optionally, one or more active ingredient(s)) uniformly throughouta solvent carrier such as water or organic solvents, such as thealcohols (e.g. ethanol or isopropanol, acetone).

In some embodiments, the solution is an aqueous solution wherein aprovided compound may be appropriately buffered by means of saline,acetate, phosphate, citrate, acetate or other buffering agents, whichmay be at any physiologically acceptable pH, generally from about pH 4to about pH 7. Combinations of buffering agents may also be employed,such as phosphate buffered saline, a saline and acetate buffer, and thelike. In the case of saline, a 0.9% saline solution may be employed. Inthe case of acetate, phosphate, citrate, acetate and the like, a 50 mMsolution may be employed. In addition to buffering agents, suitablepreservatives may be employed, to prevent or limit bacteria and othermicrobial growth. One such preservative that may be employed is 0.05%benzalkonium chloride.

In some embodiments, inventive compositions comprising an isoprenylcompound, a carrier and other, optional ingredients are provided in theform of an emulsion. Emulsions are a two-phase system prepared bycombining two immiscible liquid carriers, one of which is disburseduniformly throughout the other and consists of globules that havediameters equal to or greater than those of the largest colloidalparticles. The globule size is critical and must be such that the systemachieves maximum stability. Usually, separation of two phases will notoccur unless a third substance, an emulsifying agent, is incorporated.Thus, a basic emulsion in the context of the present invention typicallycontains two or more components (e.g., two immiscible liquid carriers,an emulsifying agent, and an isoprenyl compound). In some embodiments aprenyl compound can be an emulsifying agent. Typically, emulsionsincorporate an aqueous phase into a non-aqueous phase (or vice versa).However, it is possible to prepare emulsions that are largelynon-aqueous, for example, anionic and cationic surfactants of thenon-aqueous immiscible system glycerin and olive oil. Exemplaryemulsifying agents are described herein.

In some embodiments compositions of the present invention comprise anemulsion including AFC. In some embodiments, non-lipid-based vehiclesare useful in an emulsion comprising AFC due to the lipophilic nature ofAFC.

In some embodiments, inventive compositions comprising an isoprenylcompound, are provided in the form of gel suspensions, (a semi-solidcarrier) or solid carrier to form a paste, powder, ointment, cream,lotion, hydrogel or the like. Exemplary ointments that may be preparedas a gel-suspension include semi-solid preparations intended forexternal application to the epithelium. Generally, ointment bases arecategorized into hydrocarbon bases (oleaginous), which may use whitepetroleum as a base; adsorption bases (anhydrous), which might usehydrophilic petroleum or anhydrous lanolin; emulsion bases (water andoil type); emulsion bases (oil and water type); and water soluble bases,which often use polyethylene glycol as an ointment base.

Additional isoprenyl compositions of the present invention can bereadily prepared using technology known in the art as described inRemington's Pharmaceutical Sciences, 18^(th) or 19^(th) editions,published by the Mack Publishing Company of Easton, Pa.

It is also possible and contemplated that provided compounds of thepresent invention may be in a dried and particulate form. In certainembodiments, particles are between about 0.5 and 6.0 μm, such that theparticles have sufficient mass to settle on the lung surface, and not beexhaled, but are small enough that they are not deposited on surfaces ofthe air passages prior to reaching the lung. Any of a variety ofdifferent techniques may be used to make dry powder microparticles,including but not limited to micro-milling, spray drying and a quickfreeze aerosol followed by lyophilization. With micro-particles,provided compounds may be deposited to the deep lung, thereby providingquick and efficient absorption into the bloodstream. Further, with suchapproach penetration enhancers are not required, as is sometimes thecase in transdermal, nasal or oral mucosal delivery routes. Any of avariety of inhalers can be employed, including propellant-basedaerosols, nebulizers, single dose dry powder inhalers and multidose drypowder inhalers. Common devices in current use include metered doseinhalers, which are used to deliver medications for the treatment ofasthma, chronic obstructive pulmonary disease and the like. Preferreddevices include dry powder inhalers, designed to form a cloud or aerosolof fine powder with a particle size that is always less than about 6.0μm.

Microparticle size, including mean size distribution, may be controlledby means of the method of making. For micro-milling, the size of themilling head, speed of the rotor, time of processing and the likecontrol the microparticle size. For spray drying, the nozzle size, flowrate, dryer heat and the like control the microparticle size. For makingby means of quick freeze aerosol followed by lyophilization, the nozzlesize, flow rate, concentration of aerosoled solution and the likecontrol the microparticle size. These parameters and others may beemployed to control the microparticle size.

In some embodiments, provided compounds of the present invention may betherapeutically administered by means of an injection, typically a deepintramuscular injection, such as in the gluteal or deltoid muscle, of atime release injectable formulation. In some embodiments, providedcompounds of the present invention are formulated with a PEG, such aspoly(ethylene glycol) 3350, and optionally one or more additionalexcipients and preservatives, including but not limited to excipientssuch as salts, polysorbate 80, sodium hydroxide or hydrochloric acid toadjust pH, and the like. In some embodiments, a provided compound of thepresent invention is formulated with a poly(ortho ester), which may bean auto-catalyzed poly(ortho ester) with any of a variable percentage oflactic acid in the polymeric backbone, and optionally one or moreadditional excipients. In one embodiment poly (D,L-lactide-co-glycolide)polymer (PLGA polymer) is employed, preferably a PLGA polymer with ahydrophilic end group, such as PLGA RG502H from Boehringer Ingelheim,Inc. (Ingelheim, Germany).

Such formulations may be made, for example, by combining a providedcompound of the present invention in a suitable solvent, such asmethanol, with a solution of PLGA in methylene chloride, and addingthereto a continuous phase solution of polyvinyl alcohol under suitablemixing conditions in a reactor. In general, any of a number ofinjectable and biodegradable polymers, which are preferably alsoadhesive polymers, may be employed in a time release injectableformulation. The teachings of U.S. Pat. Nos. 4,938,763, 6,432,438, and6,673,767, and the biodegradable polymers and methods of formulationdisclosed therein, are incorporated herein by reference. The formulationmay be such that an injection is required on a weekly, monthly or otherperiodic basis, depending on the concentration and amount of a providedcompound, the biodegradation rate of the polymer, and other factorsknown to those of skill in the art.

In some embodiments, inventive compositions formulated as aqueoussuspensions wherein a provided compound is in admixture with excipientsadditives and/or suitable for the manufacture of aqueous suspensions.Such additives and/or excipients are suspending agents, for example,sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gumtragacanth, and gum acacia; dispersing or wetting agents may be anaturally occurring phosphatide such as lecithin, or condensationproducts of an alkylene oxide with fatty acids, for example,polyoxyethylene stearate, or condensation products of ethylene oxidewith long chain aliphatic alcohols, for example, heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partialesters derived from fatty acids and a hexitol such as polyoxyethylenesorbitol monooleate, or condensation products of ethylene oxide withpartial esters derived from fatty acids and hexitol anhydrides, forexample polyethylene sorbitan monooleate. Aqueous suspensions also maycontain one or more coloring agents, one or more flavoring agents, andone or more sweetening agents, such as sucrose or saccharin.

In some embodiments, inventive compositions formulated as oilysuspensions by suspending a provided compound in a vegetable oil (e.g.,arachis oil, olive oil, sesame oil, coconut oil, or a mineral oil, suchas liquid paraffin). Oily suspensions may contain a thickening agent(e.g., beeswax, hard paraffin or cetyl alcohol). Sweetening agents, suchas those described herein, and flavoring agents may be added to providea palatable oral composition. Such compositions may be preserved by theaddition of an antioxidant (e.g., ascorbic acid).

In some embodiments, inventive compositions formulated as dispersiblepowders and/or granules are suitable for compositions of an aqueoussuspension by adding water. Provided compound in such powders andgranules is provided in admixture with a dispersing or wetting agent,suspending agent, and one or more preservatives. Suitable dispersing orwetting agents and suspending agents are exemplified by those alreadymentioned herein. Additional excipients, for example, sweetening,flavoring and coloring agents also may be present.

Compositions of the invention also may be in the form of oil in wateremulsions. The oily phase may be a vegetable oil, for example, olive oilor arachis oil, or a mineral oil, for example a liquid paraffin, or amixture thereof. Suitable emulsifying agents may be naturally occurringgums, for example, gum acacia or gum tragacanth, naturally-occurringphosphatides, for example soy bean, lecithin, and esters or partialesters derived from fatty acids and hexitol anhydrides, for examplesorbitan monooleate, and condensation products of the partial esterswith ethylene oxide, for example, polyoxyethylene sorbitan monooleate.The emulsions also may contain sweetening and flavoring agents.

Compositions of the invention also may be formulated as syrups andelixirs. Syrups and elixirs may be formulated with sweetening agents,for example, glycerol, propylene glycol, sorbitol or sucrose. Suchformulations also may contain a demulcent, a preservative, and flavoringand coloring agents. Demulcents are protective agents employed primarilyto alleviate irritation, particularly mucous membranes or abradedtissues. A number of chemical substances possess demulcent properties.These substances include the alginates, mucilages, gums, dextrins,starches, certain sugars, and polymeric polyhydric glycols. Othersinclude acacia, agar, benzoin, carbomer, gelatin, glycerin, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose,propylene glycol, sodium alginate, tragacanth, hydrogels and the like.

4. Administration and Dosage Forms

Provided compounds of the invention of this invention may be formulatedby any means known in the art, including but not limited to formulationas tablets, capsules, caplets, suspensions, powders, lyophilizedpreparations, suppositories, ocular drops, skin patches, oral solubleformulations, sprays, aerosols and the like, and may be mixed andformulated with buffers, binders, excipients, stabilizers, anti-oxidantsand other agents known in the art. In general, any route ofadministration by which provided compounds of the invention areintroduced across an epidermal layer of cells may be employed.Administration means may thus include administration through mucousmembranes, buccal administration, oral administration, dermaladministration, inhalation administration, pulmonary administration,nasal administration, urethral administration, vaginal administration,and the like.

In general, compositions comprising a therapeutically orpharmaceutically effective amount of an inventive composition ofprovided compounds may be formulated for administration in unit dosageforms.

Oral Administration

Because of their ease of administration, tablets and capsules representan advantageous oral unit dosage form. If desired, a compositionincluding provided compound of the invention may be coated by standardaqueous or nonaqueous techniques. The amount of active compound, i.e.isoprenyl compounds of the present invention, in such therapeuticallyuseful compositions is such that an effective dosage will be obtained.In another advantageous dosage unit form, sublingual pharmaceuticalcompositions may be employed, such as sheets, wafers, tablets or thelike. An active compound can also be administered intranasally as, forexample, by liquid drops or spray.

The tablets, pills, capsules, and the like may also contain a binder asdescribed herein. In some embodiments, binders that may be particularlyuseful for tablets, pills and capsules, include gum tragacanth, acacia,corn starch or gelatin; excipients such as dicalcium phosphate; adisintegrating agent such as corn starch, potato starch or alginic acid;a lubricant such as magnesium stearate; and a sweetening agent such assucrose, lactose or saccharin. When a dosage unit form is a capsule, itmay contain, in addition to materials of the above type, a liquidcarrier, such as a fatty oil.

Compositions of the present invention may be in additional formssuitable for oral use, for example, troches, lozenges, pills, aqueous oroily suspensions, solutions, dispersible powders or granules, emulsions,hard or soft capsules, syrups or elixirs, pastes, gels or the like.

A tablet may contain the active ingredient(s) in admixture withnon-toxic pharmaceutically acceptable additives and/or excipients whichare suitable for the manufacture of tablets. These additives orexcipients may be, for example, fillers, wetting agents, inert diluents,such as calcium carbonate, sodium carbonate, lactose, calcium phosphateor sodium phosphate; granulating and noneffervescent disintegratingagents, (e.g., corn starch or alginic acid); binding agents (e.g.,starch, gelatin or acacia); and lubricating agents (e.g., magnesiumstearate, stearic acid or talc).

A tablet may be prepared by traditional methods such as by compressingor molding a powder or granules containing a provided compound.Compressed tablets may be prepared by compressing, in a suitablemachine, the a provided compound in a free-flowing form, such as apowder or granules optionally mixed with a binder, lubricant, inertdiluent, and/or surface active/dispersing agent(s). Molded tablets maybe made by molding, in a suitable machine, a powdered provided compoundmoistened with an inert liquid binder.

The tablets may be uncoated or they may be coated by known techniques todelay disintegration and absorption in the gastrointestinal tract andthereby provide a sustained action over a longer period. For example, atime delay material such as glyceryl monostearate or glyceryl distearatemay be employed. They also may be coated for controlled delivery. Forexample, a “delayed release” dosage form releases a product or substanceat a time other than promptly after administration. Examples ofdelayed-release systems include repeat action tablets and capsules, andenteric coated tablets where timed release is achieved by a barriercoating.

Compositions of the present invention also may be formulated for oraluse as hard gelatin capsules, where a provided compound is mixed with aninert solid diluent, for example, calcium carbonate, calcium phosphateor kaolin, or soft gelatin capsules wherein the active ingredient(s) is(are) mixed with water or an oil medium, for example, peanut oil, liquidparaffin, or olive oil.

In some embodiments, liquid preparations for oral administration canalso be used. Liquid preparations can be in the form of solutions,syrups or suspensions, or a dry product for reconstitution with water oranother suitable vehicle before use. Such liquid preparations can beprepared by conventional means with pharmaceutically acceptableadditives such as suspending agents, emulsifying agents, non-aqueousvehicles, and preservatives.

Liquid based oral dosage forms, like their solid counterparts, usuallycontain at least 0.1 mg of a provided compound. One skilled in the artwill be able to properly formulate a liquid formulation containing anappropriate amount of a provided compound per fluidic ounce, dependingon the additive or carrier selected.

Compositions intended for oral use may be prepared according to anyknown method, and such compositions may contain one or more excipientsselected from the group consisting of sweetening agents, flavoringagents, coloring agents, and preserving agents in order to providepharmaceutically elegant and palatable compositions. In general, theformulations for oral administration are prepared by uniformly andintimately admixing the active compound, i.e., a provided compound ofthe present invention or mixtures thereof, with a liquid or finelydivided solid excipient, or both, and then, if necessary, shaping theresulting mixture.

Parenteral Administration

Provided compounds of the present invention may also be administeredparenterally. Solutions or suspensions of these active peptides may beprepared in water suitably mixed with a surfactant, such ashydroxy-propylcellulose. Dispersions may also be prepared, such asdispersions in glycerol, liquid polyethylene glycols and mixturesthereof in oils. These preparations may optionally contain apreservative to prevent the growth of microorganisms. Lyophilized singleunit formulations may also be utilized, which are reconstituted, such aswith saline, immediately prior to administration, and thus do notrequire a preservative.

Pharmaceutical forms suitable for injectable use include, for example,sterile aqueous solutions or dispersions and sterile powders, such aslyophilized formulations, for the extemporaneous preparation of sterileinjectable solutions or dispersions. In all cases, the form must besterile and must be fluid to the extent that it may be administered bysyringe. The form must be stable under the conditions of manufacture andstorage and may be preserved against the contaminating action ofmicroorganisms such as bacteria and fungi. The carrier can be a solventor dispersion medium containing, for example, water, ethanol, a polyol,for example glycerol, propylene glycol or liquid polyethylene glycol,suitable mixtures thereof, and vegetable oils.

Parenterally administered compositions are formulated to allow forinjection, either as a bolus or as a continuous infusion. For parenteralapplication, “parenteral” meaning subcutaneous injections, intravenous,intramuscular, intrasternal injection, or infusion techniques,particularly suitable vehicles consist of solutions, preferably oily oraqueous solutions, as well as suspensions, emulsions, or implants.Formulations for injection can be prepared in unit dosage forms, such asampules, or in multi-dose units, with added preservatives. Thecompositions for injection can be in the form of suspensions, solutions,or emulsions, containing either oily or aqueous additives. They may alsocontain formulatory agents such as suspending agents, stabilizingagents, and/or dispersing agents. A isoprenyl compound may also bepresented in powder form for reconstitution with a suitable vehiclebefore use.

The compositions of the present invention also may be in the form of asterile injectable aqueous or oleaginous suspension. Injectablecompositions, such as sterile injectable aqueous or oleaginoussuspensions, may be formulated according to the known art using suitabledispersing or wetting agents and suspending agents. The sterileinjectable composition may also be a sterile injectable solution orsuspension in a nontoxic parenterally acceptable diluent or solvent, forexample, as a solution in 1,3 butanediol. Among the acceptable vehiclesand solvents that may be employed are water, Ringer's solution, andisotonic sodium chloride solution. In some embodiments, formulations ofthe present invention suitable for parenteral administrationconveniently comprise sterile aqueous preparations of the activecompound, i.e. a isoprenyl compound, which preparations are preferablyisotonic with the blood of the intended recipient. Such preparations mayconveniently be prepared by admixing the active compound with water or aglycine buffer and rendering the resulting solution sterile and isotonicwith the blood.

In addition, sterile, fixed oils are conventionally employed as asolvent or suspending medium. Aqueous suspensions may contain substanceswhich increase the viscosity of the suspension and include, for example,sodium carboxymethyl cellulose, sorbitol and/or dextran. Optionally, thesuspension may also contain stabilizers. Alternately, a compound of thepresent invention can be added to a parenteral lipid solution.

Buccal Administration

Formulations suitable for buccal administration include tablets andlozenges comprising a isoprenyl compound in a flavored base, such assucrose, acacia or tragacanth; and pastilles comprising the isoprenylcompound in an inert base, such as gelatin and glycerin or sucrose andacacia.

Topical Administration

Formulations of the present invention suitable for topical applicationto the skin take the form of an ointment, cream, lotion, paste, gel,spray, aerosol, or oil. Additives which may be used include vaseline,lanoline, polyethylene glycols, alcohols, transdermal enhancers, andcombinations of two or more thereof.

In some embodiments, formulations suitable for topical applicationachieve transdermal delivery. Transdermal pharmaceutical devices includepatches, occlusive dressings, occlusive formulations, hypodermic sprays,iontophoretic systems, gels and infusion pumps, all of which are wellknown in the art. A transdermal patch which includes a pharmaceuticalmay generally include a backing layer impermeable to the pharmaceutical,a reservoir to house the pharmaceutical, and an adhesive cover to beremoved upon use of the patch and for adhesion to the skin of a patient.

Formulations suitable for transdermal administration may also bepresented as medicated bandages or discrete patches adapted to remain inintimate contact with the epidermis of the recipient for a prolongedperiod of time. Representative examples of suitable transdermal patchesinclude, for example, those developed by NeuroDerm Ltd (Israel) and/orthat used to deliver estradiol, for example, those developed by NovogynePharmaceuticals. Formulations suitable for transdermal administrationmay also be delivered by iontophoresis (passage of a small electriccurrent (^(˜)15 mA) to “inject” electrically charged ions into the skin)through the skin. For this, the dosage form typically takes the form ofan optionally buffered aqueous solution of the active compound, i.e. aisoprenyl compound.

Formulations suitable for transdermal administration may also bedelivered by using an infusion pump connected to a needle that isinserted through the skin, for example, those developed by Medtronicused to deliver insulin. Amounts of compound used in a transdermaldevice as described herein may vary, depending on many factors includingthe size of the device and its release characteristics, the amount ofthe pharmaceutical active agent and the estimated duration of action ofthe device. Broadly, amounts of compound typically range from about 0.1%to about 10% w/v.

Administration by Inhalation

For administration by inhalation, compositions for use in the presentinvention can be delivered in the form of an aerosol spray in apressurized package or as a nebulizer, with use of suitable propellants.In the case of a pressurized aerosol, the dosage unit can be determinedby providing a valve to deliver a metered dose in accordance with theinvention.

5. Dosage Therapeutically Effective Amount

The actual quantity of compounds administered to a patient will varydepending on the severity and type of indication, the mode ofadministration, the particular compound used, the formulation used, andthe response desired.

The dosage for treatment is administration, by any of the foregoingmeans or any other means known in the art, of an amount sufficient tobring about the desired therapeutic effect. Thus, a therapeuticallyeffective amount may be an amount of a compound or pharmaceuticalcomposition that is sufficient to induce a desired effect, including butnot limited to an anti-inflammation effect. Those of ordinary skill inthe art will appreciate that a therapeutically effective amount may beadministered by means of a single dose or multiple doses, and thatcompositions provided herein may contain a unit dose of atherapeutically effective amount.

In general, provided compounds are highly active. For example, acompound may be administered at about 10 μg/kg to about 100 mg/kg bodyweight, depending on the specific compound selected, the desiredtherapeutic response, the route of administration, the formulation andother factors known to those of skill in the art.

5. Dosage Therapeutically Effective Amount

The actual quantity of compounds administered to a patient will varydepending on the severity and type of indication, the mode ofadministration, the particular compound used, the formulation used, andthe response desired.

The dosage for treatment is administration, by any of the foregoingmeans or any other means known in the art, of an amount sufficient tobring about the desired therapeutic effect. Thus, a therapeuticallyeffective amount may be an amount of a compound or pharmaceuticalcomposition that is sufficient to induce a desired effect, including butnot limited to an anti-inflammation effect. Those of ordinary skill inthe art will appreciate that a therapeutically effective amount may beadministered by means of a single dose or multiple doses, and thatcompositions provided herein may contain a unit dose of atherapeutically effective amount.

In general, provided compounds are highly active. For example, acompound may be administered from about 0.001 mg/kg to about 100 mg/kg,from about 0.01 mg/kg to about 50 mg/kg, from about 0.1 mg/kg to about40 mg/kg, from about 0.5 mg/kg to about 30 mg/kg, from about 0.01 mg/kgto about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, or from about1 mg/kg to about 25 mg/kg of a therapeutic agent per subject body weightper day to obtain a desired therapeutic effect. A desired dosage may bedelivered to a subject only once. A desired dosage may be delivered morethan three times per day, three times per day, two times per day, onceper day, every other day, every third day, every week, every two weeks,every three weeks, every four weeks, every two months, every six months,every twelve months, every two years, every three years, every fouryears, every five years, every 10 years, or every 20 years. In certainembodiments, the desired dosage may be delivered using multipleadministrations (e.g., two, three, four, five, six, seven, eight, nine,ten, eleven, twelve, thirteen, fourteen, fifteen, or moreadministrations). The skilled artisan will appreciate that certainfactors can influence the dosage and timing required to effectivelytreat a subject, including but not limited to the specific compoundselected, the desired therapeutic response, the route of administration,the formulation, the severity of the disease or disorder, previoustreatments, the general health and/or age of the subject, other diseasespresent, and/or other factors known to those of skill in the art.

6. Uses

In certain embodiments, the present invention provides novel isoprenylcompounds, which might themselves be added to or combined with otherpharmaceutically active agents, compositions comprising at least oneisoprenyl compounds or combination with other pharmaceutically activeagents thereof, and/or methods of their preparation or use in theamelioration, treatment or prevention of, for example, certainconditions, diseases or disorders associated with inflammation or thesuppression of inflammatory responses.

In certain particular embodiments, the present invention providesanti-inflammatory compounds and compositions described here that inhibitinflammation and are therefore useful in the treatment of diseases,conditions or disorders associated with inflammation. In certainparticular embodiments, the present invention provides pro-inflammatorycompounds and compositions described herein that promote inflammationand are therefore useful in the treatment of diseases, conditions ordisorders associated with the suppression of the inflammatory responses.

In certain embodiments, the present invention provides novel compoundsand compositions that modulate inflammation. Although not wishing to bebound by one theory, it is believed that compounds and compositionsdescribed herein modulate levels of inflammatory mediators, for example,cytokines. Non-limiting examples of inflammatory mediators modulated byprovided compounds and compositions include but are not limited toIL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10,IL-11, IL-12/IL-23 p40, IL13, IL-17, IL-18, TGF-β, IFN-γ, GM-CSF, Groα,MCP-1 and TNF-α. Although not wishing to be bound by one theory, it isbelieved that compounds and compositions described herein modulatelevels of inflammatory mediators that are associated with a variety ofsignal transduction pathways. Non-limiting examples of signaltransduction pathways that result in release of inflammatory mediatorssuch as cytokines, include but are not limited to G-protein-mediated,PPAR-mediated, Toll-like receptor-mediated, and TNF-α receptor-mediated.Although not wishing to be bound by one theory, it is believed thatprovided compounds and composition modulate T-helper cell infiltrationand accumulation. Although not wishing to be bound by one theory, it isbelieved that provided compounds and compositions inhibit oxidativeburst from neutrophils and are therefore anti-oxidants.

In certain embodiments, the present invention provides novel compoundsand compositions that relate to treating or lessening the severity ofone or more diseases in which protein inhibitors that modulate theG-protein signaling cascade are known to play a role. Although notwishing to be bound by one theory, it is believed that compounds andcompositions described herein inhibit methylesterification reactions bya specific membrane associated S-adenosylmethionine-dependentisoprenyl-S-isoprenyl methyltransferase (“ICMT”) resulting incarboxy-terminal polyisoprenoid cysteine modifications of a number ofkey factors in G-protein signaling pathway. In certain embodiments,provided compounds and compositions alter the interactions amongpolyisoprenylated signal transduction proteins, such as G-proteins andthe protein regulatory targets with which they interact, or otherintracellular signaling proteins.

In certain embodiments, such compounds are administered in vitro. Incertain embodiments such compounds are administered in vivo.

Another aspect of the present invention is directed to methods oftreating, preventing, or ameliorating inflammation by administering aneffective amount of a provided compound.

In some embodiments, one or more inventive compounds, alone or togetherwith one or more other pharmaceutically active agents, is used to whitenskin. In some such embodiments, an isoprenyl compound is appliedtopically.

In general, the actual quantity of provided compounds of the inventionadministered to a patient will vary depending on the severity and typeof indication, the mode of administration, the particular compound used,the formulation used, and the response desired.

The dosage for treatment is administration, by any of the foregoingmeans or any other means known in the art, of an amount sufficient tobring about the desired therapeutic effect. Thus, an effective amountincludes an amount of a provided compound (or mixture of providedcompounds) or pharmaceutical composition of this invention that issufficient to induce a desired effect, including specifically ananti-inflammation effect or a proinflammatory effect depending on thediseases, disorders, conditions, syndromes, and the like, being treated,prevented or promoted.

In general, the provided compounds of the present invention are highlyactive. For example, a provided compound can be administered at about 10μg/kg to about 50 mg/kg body weight, depending on the specific providedcompound selected, the desired therapeutic response, the route ofadministration, the formulation and other factors known to those ofskill in the art.

Methods (A) Antiinflammatory

Specifically, the present invention relates to a method of treating orlessening the severity of inflammatory diseases or disorders selectedfrom inflammation (acute or chronic), inflammatory diseases or disorders(e.g., asthma, autoimmune diseases, and COPD including emphysema,chronic bronchitis and small airways disease, etc.), inflammatoryresponses of the immune system, skin diseases (e.g., reducing acute skinirritation for patients suffering from rosacea, atopic dermatitis,seborrheic dermatitis, psoriasis), irritable bowel syndrome (e.g.,Chron's disease and ulcerative colitis, etc.), NeurodegenerativeDisorders (Parkinson's disease, Alzheimer's disease, Huntington'sdisease, Dementia pugilistica, Pick's disease, Guam parkinsonismdementia complex, Fronto-temporal dementia, Cortico-basal degeneration,Pallido-pontal-nigral degeneration, Progressive supranuclear palsy,Dementia with Lewy bodies (DLB), and multiple system atrophy (MSA)), aswell as inflammation associated with spinal cord injury to promote nerveregeneration and inhibition of rejection of genetically engineered cellsby the immune sustem during in vivo gene therapy, wherein the methodcomprises administering to a patient in need thereof a composition ofthe present invention.

In some embodiments, the provided compounds of the present invention arecapable of effectively inhibiting inflammatory responses. Thus, providedcompounds are inhibitors of edema, erythema and myeloperoxidase and aretherefore useful for treating one or more disorders associated withinflammatory diseases or disorders as described herein. In particular,the present invention encompasses the finding that certain compoundshaving superior in vivo activity than other compounds in the same class.For example, relative to AFC, compound A has improved edema inhibition,improved erythema inhibition and improved MPO (myeloperoxidase)inhibition. Therefore, such compounds are administered to a subjectsuffering from or susceptible to one or more inflammatory diseases ordisorders.

In some embodiments, the provided anti-inflammatory compounds of thepresent invention are capable of effectively inhiting inflammatoryresponses by decreasing the levels or production of inflammatorymediators such as inflammatory cytokines, for example TNF IL-1α, IL-1β,IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11,IL-12/IL-23 p40, IL13, IL-17, IL-18, TGF-β, IFN-γ, GM-CSF, Groα, MCP-1and TNF-α. Thus, provided anti-inflammatory compounds are inhibitors ofproinflammatory cytokines and are therefore useful in treating one ormore disorders associated with inflammatory diseases, conditions ordisorders described herein. In particular, the present inventionencompasses the finding that certain compounds have superior activity,as measured by percent inhibition of levels or production ofproinflammatory cytokines in animal and cell-based inflammatory models,than other compounds in the same class. Therefore, such compounds areadministered to a subject suffering from or susceptible to one or moreinflammatory diseases, conditions or diseases.

In some embodiments, the treatment of inflammatory diseases or disordersis achieved using compounds without having the side effects ofcorticosteroids or NSAIDS.

In some embodiments, the provided compounds of the present invention arecapable of effective inhibiting oxidative burst response fromneutrophils. Thus, provided compounds are inhibitors of oxidative burstresponse and are therefore useful in the treatment or amelioration ofsymptoms relating to oxidative damage caused by chemical orenvironmental factor (e.g., UV damage on the skin). In particular, thepresent invention encompasses the finding that certain compounds havesuperior activity, as measured by percent reduction in superoxideformation, than other compounds in the same class. Therefore, suchcompounds are administered to a subject suffering from conditionsassociated with oxidative damage. In some embodiments, combinations ofsuch sun screening agents with isoprenyl compounds provided hereinexhibit anti-oxidant effects (e.g., inhibition of superoxide formation).

(B) Immune Stimulatory

In some embodiments, certain compounds of the present invention arecapable of promoting inflammatory responses, and are thereforeproinflammatory. Thus, provided proinflammatory compounds are promotersof edema, erythema and myeloperoxidase (a marker for neutrophilinfiltration) and are therefore useful for treating one or moredisorders associated with the suppression of inflammatory responses asdescribed herein. Therefore, such compounds are administered to asubject suffering from or susceptible to one or more diseases,conditions or disorders associated with suppression of inflammatoryresponses.

In some embodiments, the present invention relates to a method oftreating or lessening the severity of diseases, conditions or disordersassociated with the suppression of inflammatory responses selected fromfor example, treatment of secondary bacterial or viral infectionsinflicting subjects with acquired immune deficiency syndrome (AIDS),suppression of systemic inflammatory response syndromes following severeburn injuries and cardiac surgeries and also the side-effect of a numberof drugs, for example thalidomide.

(C) Skin Conditions

In some embodiments, provided herein is a method for treating orpreventing a skin condition, the method comprising the step of topicallyapplying onto a surface of a subject, including a human, in needthereof, an effective amount of a composition comprising at least oneisoprenyl compound, a carrier and optionally an additional activeingredient. In another aspect, provided herein is a method for treatingor preventing a skin condition, the method comprising the step oftopically applying onto a surface of a subject, including a human, inneed thereof, at least 0.1 mg of a compound of Formula I. In a furtherembodiment, provided herein is a method of promoting healthy skin in asubject, including a human, in need thereof, the method comprising thestep of topically applying onto a surface of a subject, including ahuman, in need thereof, an effective amount of a composition comprisingat least one isoprenyl compound, a carrier and optionally an additionalactive ingredient. In a further aspect, provided herein is a method ofpromoting healthy skin in a subject, including a human, in need thereof,the method comprising the step of topically applying onto a surface of asubject, including a human, in need thereof, at least 0.1 mg of acompound of Formula I′.

In a further embodiment, the present invention provides a method fortreating or preventing inflammation in a subject, including a human, inneed thereof, comprising the step of administering an effective amountof a composition comprising at least one isoprenyl compound, a carrierand optionally an additional active ingredient. In a further aspect, thepresent invention provides a method for treating or preventinginflammation in a subject, including a human, in need thereof,comprising the step of administering at least 0.1 mg of a compound ofFormula I′.

In certain embodiments, the present invention provides uses of providedcompounds and/or compositions in the treatment or prevention of diseasesor conditions associated with suppression of inflammatory responses. Incertain embodiments, the present invention provides a composition fortreating or preventing conditions associated with suppression of theinflammatory responses, in a subject, including a human, in need oftreatment thereof, that comprises of at least one isoprenyl compound, acarrier and optionally, an additional active ingredient. In a furtherembodiment, provided herein is a method for treating or preventing adisease or condition associated with suppression of inflammatoryresponses, in a subject, including a human, in need thereof, the methodcomprising the step of administering an effective amount of acomposition comprising at least one isoprenyl compound, a carrier andoptionally an additional active ingredient. In a further aspect,provided herein is a method for treating or preventing a disease orcondition associated with suppression of inflammatory responses, in asubject, including a human, in need thereof, the method comprising thestep of administering at least 0.1 mg of a compound of Formula I′.

Exemplary diseases, disorders or conditions (e.g., rosacea, psoriasis,atopic dermatitis and seborrhic dermatitis) that may be treated withcompounds of Formulae I, I′, and/or or Ia, in accordance with thepresent invention are addressed individually below.

Rosacea

Rosacea is a chronic, inflammatory skin disorder that afflicts about 14million people in the US (FoxAnalytics, The Dermatology Market Outlookto 2011, B.I. LTD, Editor: London, UK, p. 201; Crandall, M. A. MarketIntelligence Report, K. Information, Editor, 2008: New York. p 359).With peak onset between the ages of 51 and 60, its incidence will growsubstantially in the years ahead. The condition is characterized by aconstellation of symptoms that include central facial erythema,telangiectasias, papules, granulomatous nodules, phyma formation andocular changes. Flares and remissions occur without rationale. There areno known cures for rosacea. Exemplary cytokines associated with rosaceamay include TNFα, ILβ, IL-6, IL-8, MCP-1 and Groα.

Psoriasis

Psoriasis is a chronic inflammatory skin disease affecting ˜125 millionpeople worldwide and approximately 2-3% of the general population in theUS and Europe (Crandall, M. A. Market Intelligence Report, K.Information, Editor, 2008: New York. P. 359; Naldi, L., Curr. DrugTargets Inflamm. Allergy, 2004, 3: 121-128). Although the pathogenesisof psoriasis has not been fully elucidated, recent advances demonstratetargeting key mediators of inflammation as a promising therapeuticapproach (Numerof et al., BioDrugs, 2006, 20: 93-103; Menter et al., J.Am. Acad. Dermatol., 2009, 60: 643-659). Direct therapeutic approachesinclude using antibodies or soluble receptors (i.e., biologics) todirectly neutralize the specific cytokine of interest. However, biologiccytokine-derived therapies are expensive to produce, require sustainedhigh blood levels in order to develop significant skin levels, mayinduce the production of neutralizing antibodies (leading to adiminished response to therapy), and must be administered by injection.Topical treatments have largely been ineffective, so market growth hasbeen driven by systemic agents that have serious potential side effects.Corticosteroids remain the cornerstone of current topical treatment, butthey are far from ideal. Long-term steroid use brings safety concernsranging from issues of systemic absorption to cutaneous atrophy and itsvarious clinical presentations. Today's US market for psoriasistreatments is greatly underserved, as only 60% of sufferers are beingtreated (Horn et al., J. Am. Acad. Dermatol. 2007, 57: 957-962).

Psoriasis can be conceived in simple terms, as a self reinforcing loop,in which deregulated inflammatory activity stimulates the epidermalStat3c signaling pathway in the epidermis resulting in epidermalhyperplasia. The affected keratinocytes secrete cytokines which simulatethe immune system, including T-helper cell (THc) infiltration andaccumulation. Cytokines from the activated immune cells positivelyfeedback on to the epidermal Stat3c pathway maintaining and amplifyingthe pathophysiology. Inhibition of THc infiltration and accumulationwould decrease Stat3c expression and the onset of psoriasis. Exemplarycytokines associated with psoriasis may include TNFα, IL1α, IL1β, IL-2,IL-6, IL-8, IL-12, MCP-1, Groα and IFNγ.

In some embodiments, compounds of the present invention show asurprising inhibition of T-helper cell infiltration and accumulation.

Inflammatory Cytokines and Psoriasis

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditionssuch as psoriasis comprising administering to a subject in need thereofa dosage form comprising at least about 0.1 mg of a isoprenyl compoundof Formulae I, I′ and/or in described classes and subclasses thereof,wherein cytokine levels and/or activity (e.g., TNF-α levels and/oractivity) are reduced by more than about 20% (e.g., as determined usinga K5.Stat3c psoriasis mouse model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditionssuch as psoriasis comprising administering to a subject in need thereofa dosage form comprising at least about 0.1 mg of a isoprenyl compoundof Formulae I, I′ and/or in described classes and subclasses thereof,wherein cytokine levels and/or activity (e.g., IL-α levels and/oractivity) are reduced by more than about 20% (e.g., as determined usinga K5.Stat3c psoriasis mouse model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditionssuch as psoriasis comprising administering to a subject in need thereofa dosage form comprising at least about 0.1 mg of a isoprenyl compoundof Formulae I, I′ and/or in described classes and subclasses thereof,wherein cytokine levels and/or activity (e.g., IL-β levels and/oractivity) are reduced by more than about 20% (e.g., as determined usinga K5.Stat3c psoriasis mouse model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditionssuch as psoriasis comprising administering to a subject in need thereofa dosage form comprising at least about 0.1 mg of a isoprenyl compoundof Formulae I, I′ and/or in described classes and subclasses thereof,wherein cytokine levels and/or activity (e.g., IL-2 levels and/oractivity) are reduced by more than about 20% (e.g., as determined usinga K5.Stat3c psoriasis mouse model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditionssuch as psoriasis comprising administering to a subject in need thereofa dosage form comprising at least about 0.1 mg of a isoprenyl compoundof Formulae I, I′ and/or in described classes and subclasses thereof,wherein cytokine levels and/or activity (e.g., IL-6 levels and/oractivity) are reduced by more than about 20% (e.g., as determined usinga K5.Stat3c psoriasis mouse model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditionssuch as psoriasis comprising administering to a subject in need thereofa dosage form comprising at least about 0.1 mg of a isoprenyl compoundof Formulae I, I′ and/or in described classes and subclasses thereof,wherein cytokine levels and/or activity (e.g., IL-8 levels and/oractivity) are reduced by more than about 20% (e.g., as determined usinga K5.Stat3c psoriasis mouse model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditionssuch as psoriasis comprising administering to a subject in need thereofa dosage form comprising at least about 0.1 mg of a isoprenyl compoundof Formulae I, I′ and/or in described classes and subclasses thereof,wherein cytokine levels and/or activity (e.g., IL-12 levels and/oractivity) are reduced by more than about 20% (e.g., as determined usinga K5.Stat3c psoriasis mouse model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditionssuch as psoriasis comprising administering to a subject in need thereofa dosage form comprising at least about 0.1 mg of a isoprenyl compoundof Formulae I, I′ and/or in described classes and subclasses thereof,wherein cytokine levels and/or activity (e.g., IFN-γ levels and/oractivity) are reduced by more than about 20% (e.g., as determined usinga K5.Stat3c psoriasis mouse model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditionssuch as psoriasis comprising administering to a subject in need thereofa dosage form comprising at least about 0.1 mg of a isoprenyl compoundof Formulae I, I′ and/or in described classes and subclasses thereof,wherein cytokine levels and/or activity (e.g., MCP-1 levels and/oractivity) are reduced by more than about 20% (e.g., as determined usinga K5.Stat3c psoriasis mouse model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditionssuch as psoriasis comprising administering to a subject in need thereofa dosage form comprising at least about 0.1 mg of a isoprenyl compoundof Formulae I, I′ and/or in described classes and subclasses thereof,wherein cytokine levels and/or activity (e.g., Gro-α levels and/oractivity) are reduced by more than about 20% (e.g., as determined usinga K5.Stat3c psoriasis mouse model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditionssuch as psoriasis comprising administering to a subject in need thereofa dosage form comprising at least about 0.1 mg of a isoprenyl compoundof Formulae I, I′ and/or in described classes and subclasses thereof,having an activity in the inhibition of (more than about 20%) of levelsof CD3+ T-helper cells, determined using a K5.Stat3c psoriasis mousemodel.

Atopic Dermatitis

Atopic dermatitis, or eczema, is characterized by chromic inflammationand irritation of the skin. Its causes are varied but immunological innature. In the US, prevalence is 10% to 20% in children and 1% to 3% inadults. Topical dermatitis is caused by exposure to substances such aspoison ivy, detergents and cosmetics that trigger allergic skinreactions. According to present theories, Atopic dermatitis is thoughtto be caused by skin barrier defects that lead to increased exposure tosubstances such as allergens exposed by inhalation or ingestion. Whendermatitis occurs, corticosteroids are the primary treatment. Atopicdermatitis, however, disproportionately affects children, and long-termsteroid use in this population raises safety concerns. Exemplarycytokines associated with atopic dermatitis include but are not limitedto TNFα, IL1β, IL-6, IL-8, MCP-1, Groα, IL-4, IL-5, IL-10, IL-13, IL-17and IFNγ.

Inflammatory Cytokines and Atopic Dermatitis

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions such as atopic dermatitis comprising administering to asubject in need thereof a dosage form comprising at least about 0.1 mgof a isoprenyl compound of Formulae I, I′ and/or in described classesand subclasses thereof, wherein cytokine levels and/or activity (e.g.,TNF-α levels and/or activity) are reduced by more than about 20% (e.g.,as determined using a ovalbumin-challenged ft/ft atopic dermatitis mousemodel).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions such as atopic dermatitis comprising administering to asubject in need thereof a dosage form comprising at least about 0.1 mgof a isoprenyl compound of Formulae I, I′ and/or in described classesand subclasses thereof, wherein cytokine levels and/or activity (e.g.,IL-α levels and/or activity) are reduced by more than about 20% (e.g.,as determined using a ovalbumin-challenged ft/ft atopic dermatitis mousemodel).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions such as atopic dermatitis comprising administering to asubject in need thereof a dosage form comprising at least about 0.1 mgof a isoprenyl compound of Formulae I, I′ and/or in described classesand subclasses thereof, wherein cytokine levels and/or activity (e.g.,IL-β levels and/or activity) are reduced by more than about 20% (e.g.,as determined using a ovalbumin-challenged ft/ft atopic dermatitis mousemodel).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions such as atopic dermatitis comprising administering to asubject in need thereof a dosage form comprising at least about 0.1 mgof a isoprenyl compound of Formulae I, I′ and/or in described classesand subclasses thereof, wherein cytokine levels and/or activity (e.g.,IL-2 levels and/or activity) are reduced by more than about 20% (e.g.,as determined using a ovalbumin-challenged ft/ft atopic dermatitis mousemodel).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions such as atopic dermatitis comprising administering to asubject in need thereof a dosage form comprising at least about 0.1 mgof a isoprenyl compound of Formulae I, I′ and/or in described classesand subclasses thereof, wherein cytokine levels and/or activity (e.g.,IL-6 levels and/or activity) are reduced by more than about 20% (e.g.,as determined using a ovalbumin-challenged ft/ft atopic dermatitis mousemodel).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions such as atopic dermatitis comprising administering to asubject in need thereof a dosage form comprising at least about 0.1 mgof a isoprenyl compound of Formulae I, I′ and/or in described classesand subclasses thereof, wherein cytokine levels and/or activity (e.g.,IL-8 levels and/or activity) are reduced by more than about 20% (e.g.,as determined using a ovalbumin-challenged ft/ft atopic dermatitis mousemodel).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions such as atopic dermatitis comprising administering to asubject in need thereof a dosage form comprising at least about 0.1 mgof a isoprenyl compound of Formulae I, I′ and/or in described classesand subclasses thereof, wherein cytokine levels and/or activity (e.g.,IL-12 levels and/or activity) are reduced by more than about 20% (e.g.,as determined using a ovalbumin-challenged ft/ft atopic dermatitis mousemodel).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions such as atopic dermatitis comprising administering to asubject in need thereof a dosage form comprising at least about 0.1 mgof a isoprenyl compound of Formulae I, I′ and/or in described classesand subclasses thereof, wherein cytokine levels and/or activity (e.g.,IFN-γ levels and/or activity) are reduced by more than about 20% (e.g.,as determined using a ovalbumin-challenged ft/ft atopic dermatitis mousemodel).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions such as atopic dermatitis comprising administering to asubject in need thereof a dosage form comprising at least about 0.1 mgof a isoprenyl compound of Formulae I, I′ and/or in described classesand subclasses thereof, wherein cytokine levels and/or activity (e.g.,MCP-1 levels and/or activity) are reduced by more than about 20% (e.g.,as determined using a ovalbumin-challenged ft/ft atopic dermatitis mousemodel).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions such as atopic dermatitis comprising administering to asubject in need thereof a dosage form comprising at least about 0.1 mgof a isoprenyl compound of Formulae I, I′ and/or in described classesand subclasses thereof, wherein cytokine levels and/or activity (e.g.,Gro-α levels and/or activity) are reduced by more than about 20% (e.g.,as determined using a ovalbumin-challenged ft/ft atopic dermatitis mousemodel).

Seborrhic Dermatitis

Seborrheic dermatitis, commonly called dandruff, is a disease thatcauses redness, itchiness, and flaking of the skin. It affects thescalp, face, trunk, and particularly the sebum-gland rich areas of theskin, usually causing the skin to look inflamed and scaly.

Seborrheic dermatitis most often occurs in adults from 30 to 60 years ofage and is more common in men than in women. Although the exact cause isnot known, those afflicted with seborrhoeic dermatitis often have anunfavorable epidermic response caused by infections. Seborrheicdermatitis has also been linked to neurologic disorders such asParkinson's disease and epilepsy. The treatment of seborrheic dermatitisdepends on its location on the body. Treatment also depends on theperson's age. Dandruff is often treated with a shampoo that containssalicylic acid, the prescription medicine selenium sulfide, zincpyrithione, ketoconazole or coal tar. Steroid lotions may be used inadolescents and adults. Exemplary cytokines associated with seborrhicdermatitis include but are not limited to TNFα, ILβ, IL-6, IL-8, MCP-1,and Groα.

Inflammatory Cytokines and Rosacea, Psoriasis, Atopic Dermatitis andSeborrhic Dermatitis

As described herein, the present invention provides methods of treatingameloriating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis).

In some embodiments, the present invention provides methods of treatingameloriating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)by administering a compound and/or composition of Formula I, providedthat at least 0.1 mg of the compound is administered. In certainembodiments, the present invention provides methods of treatingameloriating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)by administering a compound and/or composition of Formulae I, I′ and/orin described classes and subclasses thereof, provided that at least 2 mgof the compound is administered.

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions (e.g., rosacea, psoriasis, atopic dermatitis and seborrhicdermatitis) comprising administering to a subject in need thereof adosage form comprising at least about 0.1 mg of a isoprenyl compound ofFormulae I, I′ and/or in described classes and subclasses thereof,wherein inflammatory activity (e.g., MPO activity) is reduced by morethan about 30% (e.g., as determined using an MPO activity assay).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions (e.g., rosacea, psoriasis, atopic dermatitis and seborrhicdermatitis) comprising administering to a subject in need thereof adosage form comprising at least about 0.1 mg of a isoprenyl compound ofFormulae I, I′ and/or in described classes and subclasses thereof,wherein inflammatory activity (e.g., MPO activity) is reduced by morethan about 60% (e.g., as determined using an MPO activity assay).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions (e.g., rosacea, psoriasis, atopic dermatitis and seborrhicdermatitis) comprising administering to a subject in need thereof adosage form comprising at least about 0.1 mg of a isoprenyl compound ofFormulae I, I′ and/or in described classes and subclasses thereof,wherein inflammatory activity (e.g., erythema activity) is reduced bymore than about 30% (e.g., as determined using an erythema activityassay).

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing inflammatory skinconditions (e.g., rosacea, psoriasis, atopic dermatitis and seborrhicdermatitis) comprising administering to a subject in need thereof adosage form comprising at least about 0.1 mg of a isoprenyl compound ofFormulae I, I′ and/or in described classes and subclasses thereof,wherein inflammatory activity (e.g., edema activity) is reduced by morethan about 30% (e.g., as determined using an edema activity assay).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., TNF-α, levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TPA-induced mouseear inflammatory model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-1β levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TPA-induced mouseear inflammatory model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-8 levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TPA-induced mouseear inflammatory model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-6 levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TPA-induced mouseear inflammatory model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., MCP-1 levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TPA-induced mouseear inflammatory model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., Groα levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TPA-induced mouseear inflammatory model).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., TNF-α levels and/or activity) are reducedby more than about 20% (e.g., as determined using an LPS-TLR4-inducedcytokine release inflammatory model in HMEC-1 cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-1β levels and/or activity) are reducedby more than about 20% (e.g., as determined using an LPS-TLR4-inducedcytokine release inflammatory model in HMEC-1 cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-8/KC levels and/or activity) arereduced by more than about 20% (e.g., as, determined using anLPS-TLR4-induced cytokine release inflammatory model in HMEC-1 cellline).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-6 levels and/or activity) are reducedby more than about 20% (e.g., as determined using an LPS-TLR4-inducedcytokine release inflammatory model in HMEC-1 cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., MCP-1 levels and/or activity) are reducedby more than about 20% (e.g., as determined using an LPS-TLR4-inducedcytokine release inflammatory model in HMEC-1 cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., Gro-α levels and/or activity) are reducedby more than about 20% (e.g., as determined using an LPS-TLR4-inducedcytokine release inflammatory model in HMEC-1 cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., TNF-α levels and/or activity) are reducedby more than about 20% (e.g., as determined using an ATPγS-purinergicreceptor-induced cytokine release inflammatory model in HMEC-1 cellline).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-1β levels and/or activity) are reducedby more than about 20% (e.g., as determined using an ATPγS-purinergicreceptor-induced cytokine release inflammatory model in HMEC-1 cellline).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-8/KC levels and/or activity) arereduced by more than about 20% (e.g., as determined using anATPγS-purinergic receptor-induced cytokine release inflammatory model inHMEC-1 cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-6 levels and/or activity) are reducedby more than about 20% (e.g., as determined using an ATPγS-purinergicreceptor-induced cytokine release inflammatory model in HMEC-1 cellline).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., MCP-1 levels and/or activity) are reducedby more than about 20% (e.g., as determined using an ATPγS-purinergicreceptor-induced cytokine release inflammatory model in HMEC-1 cellline).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., Gro-α levels and/or activity) are reducedby more than about 20% (e.g., as determined using an ATPγS-purinergicreceptor-induced cytokine release inflammatory model in HMEC-1 cellline).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., TNF-α levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TPA-induced cytokinerelease inflammatory model in NHEK cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-1β levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TPA-induced cytokinerelease inflammatory model in NHEK cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-8/KC levels and/or activity) arereduced by more than about 20% (e.g., as determined using a TPA-inducedcytokine release inflammatory model in NHEK cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-6 levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TPA-induced cytokinerelease inflammatory model in NHEK cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., MCP-1 levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TPA-induced cytokinerelease inflammatory model in NHEK cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., Gro-α levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TPA-induced cytokinerelease inflammatory model in NHEK cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., TNF-α levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TNFα-inducedcytokine release inflammatory model in HUVEC cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-1β levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TNFα-inducedcytokine release inflammatory model in HUVEC cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity are reduced by more than about 20%, such asIL-8/KC, determined using a TNFα-induced cytokine release inflammatorymodel in HUVEC cell line.

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., IL-6 levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TNFα-inducedcytokine release inflammatory model in HUVEC cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., MCP-1 levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TNFα-inducedcytokine release inflammatory model in HUVEC cell line).

In some embodiments, the present invention provides methods of treating,ameliorating, controlling, or preventing inflammatory skin conditions(e.g., rosacea, psoriasis, atopic dermatitis and seborrhic dermatitis)comprising administering to a subject in need thereof a dosage formcomprising at least about 0.1 mg of a isoprenyl compound of Formulae I,I′ and/or in described classes and subclasses thereof, wherein cytokinelevels and/or activity (e.g., Gro-α levels and/or activity) are reducedby more than about 20% (e.g., as determined using a TNFα-inducedcytokine release inflammatory model in HUVEC cell line).

Sun Screen (Protection from UV Damage)

Oxidative stresses caused by environmental insults such as ultraviolet(“UV”) rays from the sun, cigarette smoke exposure, consumption of foodswith high saturated fat and environmental pollutants as well as thenatural process of aging, contributing to the generation of freeradicals and reactive oxygen species (“ROS”), stimulate inflammatoryresponses, especially in the skin (Pilla et al. Intl J. Cosm. Sci. 2005v27 p 17-34). High levels of ROS contribute to adverse effects on theskin including erythema, edema, photoaging and skin cancer (Trouba etal. Antioxid. Redox Signal 2002 v4 p 665-673). Neutrophil infiltrationduring inflammatory responses is associated with increased oxygenconsumption and generation of ROS. Extracellular inflammatory agonistssuch as fMLP bind to GPCRs sich as formyl peptide receptors (“FPR”), totrigger the oxidative burst response (i.e., the rapid rapid release ofROS).

In certain embodiments, the present invention provides methods oftreating, ameloriating, controlling, or preventing UV damage toespecially the skin of a subject, in need thereof, by administering acompound and/or composition of Formula I, provided that at least 0.1 mgof the compound. In certain embodiments, the present invention providesmethods of treating, ameloriating, controlling, or preventing UV damageto especially the skin of a subject, in need thereof, by administering acompound and/or composition of Formula I, provided that at least 2 mg ofthe compound is administered.

According to one aspect, the present invention provides methods oftreating, ameliorating, controlling, or preventing UV damage toespecially the skin of a subject, in need thereof, comprisingadministering to a subject in need thereof a dosage form comprising atleast about 0.1 mg of a isoprenyl compound of Formulae I, I′ and/or indescribed classes and subclasses thereof, having an activity in theinhibition of more than about 20% of superoxide formation.

7. Combination Therapy

It is contemplated that a provided compound can be used in combinationwith other drugs or therapeutic agents.

In some embodiments, isoprenyl compounds as described herein areadministered in combination with one or more other agents intended totreat the same condition, or disease. As used herein, additionaltherapeutic agents that are normally administered to treat a particulardisease, or condition, are known as “appropriate for the disease, orcondition, being treated.”

For example, in some embodiments, compounds of the present invention, ora pharmaceutically acceptable composition thereof, are administered incombination with other anti-inflammatory agents to treat inflammatorydiseases and/or disorders. Examples of known anti-inflammatory agentsinclude, but are not limited to, dexamethasone, indomethacin andclobetasol.

In some embodiments, isoprenyl compounds of the present invention areadministered in combination with one or more other pharmaceuticallyactive agents intended to treat a different disease, disorder, orcondition. For example, in some embodiments, it may be desirable toadminister an inventive compound in order to reduce inflammation whileconcurrently administering a different pharmaceutically active agent inorder to achieve a different biological result.

To give but one example, it is known that transdermal administration ofpharmaceutically active agents often causes skin irritation at the siteof delivery. Indeed, it is not uncommon that a skin irritating agent(e.g., SDS) be administered prior to or concurrent with application of atransdermal device such as, for example, a transdermal patch, in orderto facilitate the delivery. Applicants have found that addition orco-administration of a isoprenyl compound as described herein incombination with transdermal administration of another pharmaceuticallyactive agent can reduce inflammation and/or irritation associated withthe transdermal administration of the other pharmaceutically activeagent.

It is also known that single or chronic injections of a pharmaceuticallyactive agent may sometimes result in inflammation, whether due to theidentity of the pharmaceutically active agent (i.e., as an irritant) orto the mode of delivery. The present invention contemplatesco-administration of one or more compounds of the present invention, inorder to reduce inflammation associated with single or chronic injectionof a pharmaceutically active agent.

Exemplary pharmaceutically active agents whose delivery, whethertransdermally or by injection, may cause skin irritation includelevadopa, pro-drug forms of levadopa, insulin, estradiol, estrogen,progesterone, progestins, progestogen, testosterone, nicotine,nitroglycerin, cholinesterase inhibitors, stimulants, antidepressants,and analgesics.

To give another example, application of certain agents such as, forexample, hair relaxants, which commonly are or contain basic agents(e.g., NaOH), can cause skin irritation (e.g., irritation and/orinflammation of the scalp). According to the present invention, one ormore isoprenyl compounds can be administered together with such a hairrelaxant (or other agent) to reduce skin irritation and/or inflammation.

Although the invention has been described in detail with particularreference to these preferred embodiments, other embodiments can achievethe same results. Variations and modifications of the present inventionwill be obvious to those skilled in the art and it is intended to coverall such modifications and equivalents. The entire disclosures of allreferences, applications, patents, and publications cited above and/orin the attachments, and of the corresponding application(s), are herebyincorporated by reference.

EXAMPLES

As depicted in the Examples below, in certain exemplary embodiments,compounds are prepared according to the following general procedures. Itwill be appreciated that, although the general methods depict thesynthesis of certain compounds of the present invention, the followinggeneral methods, and other methods known to one of ordinary skill in theart, can be applied to all classes, subclasses and species of each ofthese compounds, disclosed herein.

The AFC compounds, including S-trans, trans-farnesyl-L-cysteine,utilized as starting materials may be synthesized according to methodsknown in the art or synthesized by the methods disclosed in Brown etal., J Am Chem Soc, 1991, 113: 3176-3177, the disclosure of which isincorporated by reference herein. Other starting materials such asS-trans, trans-farnesyl-L-cysteine methyl ester, may be synthesizedaccording to methods known in the art or synthesized by the methodsdisclosed in Troutman et al., Bioconjugate Chem, 2005, 16: 1209-1217.

The following general experimental procedures were used for Examples1-78 as described below. Proton Nuclear Magnetic Resonance (¹H-NMR)spectroscopy was recorded on a Bruker 500 MHz spectrometer, dimethylsulfoxide (DMSO-d6), methanol (CD₃OD) or chloroform (CDCl₃) was used as¹H-NMR solvent. The residual proton absorption of the deuterated solventwas used as the internal standard. All ¹H-NMR chemical shift arereported as δ values in the parts per million (ppm). The splittingpattern abbreviations are as follows: s, singlet; d, doublet; t,triplet; q, quartet; br, broad; m, multiplet; dd, doublet of doublet;dt, doublet of triplets. The HPLC analysis was done using a phenomenexluna C₁₈(2) 50×4.6 mm column. The mobile phase is 60% water, 40%acetonitrile containing 0.05% trifluoroacetic acid at 2 mL per minuteflow rate for the first 2.5 minutes, followed by a gradient to 100%acetonitrile containing 0.05% TFA over 10 minutes. The eluent isobserved at 214 nm.

Example 1

Synthesis of(4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid) (Compound B): To a solution of S-trans, trans-farnesyl-L-cysteine(500 mg, 1.54 mmol) in THF, a first portion of K₂CO₃ (2 mmol) was addedand the resultant solution was cooled to 5° C. with vigorous stirring.To this stirred solution was added succinic anhydride (308 mg, 3.1 mmol)dropwise while maintaining the pH at 9.0-10.0 with another portion ofK₂CO₃ (4 mmol). The mixture was stirred at room temperature for 2 h,HPLC analysis showed completion of the reaction. The pH of the reactionmixture then adjusted to 2.0 by the addition of 2 N HCl solution. Theacidic solution was extracted three times with 10 mL of ethyl acetate.The combined organic extract was washed with water, brine and dried overNa₂SO₄, the solvent was removed on rotary evaporator to afford crudeCompound B, which was further purified by preparative HPLC (535 mg, 82%)to yield Compound B. ¹H-NMR (500 MHz, CDCl₃): δ 1.59 (s, 6H), 1.66 (s,6H), 2.05 (m, 8H), 2.60 (m, 2H), 2.48 (m, 2H), 2.86 (dd, 1H), 2.94 (dd,1H), 3.10 (dd, 1H), 3.12 (dd, 1H), 4.68 (dd, 1H), 5.06 (m, 2H), 5.20 (t,1H). ¹³C-NMR (125 MHz, CDCl₃): δ 16.0, 16.1, 17.7, 25.7, 26.5, 26.7,29.4, 29.8, 30.5, 32.6, 39.6, 39.7, 52.2, 119.3, 123.8, 124.3, 131.3,135.4, 140.3, 173.4, 174.2, 176.8; ES-MS: mass calcd for ChemicalFormula: C₂₂H₃₅NO₅S 425.6. Found (M+Na) m/z 448.

Example 2

Synthesis of((E)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobut-2-enoicacid) (Compound A)

A solution of S-trans, trans-farnesyl-L-cysteine (500 mg, 1.54 mmol) inTHF and a first portion of K₂CO₃ (3 mmol) was cooled to 5° C. withvigorous stirring. To this stirred solution was added maleic anhydride(302 mg, 3.07 mmol) portionwise while maintaining the pH at 9.0-10.0with another portion of K₂CO₃ (3 mmol). The mixture was stirred at roomtemperature for 3 h, HPLC analysis showed completion of the reaction.The pH of the reaction mixture then adjusted to 2.0 by the addition of 2N HCl solution. The acidic solution was extracted three times with 15 mLof ethyl acetate. The combined organic extract was washed with water,brine and dried over Na₂SO₄ and then concentrated to afford crudeCompound A, which was further purified by preparative HPLC (552 mg, 85%)to yield Compound A. ¹H-NMR (500 MHz, CD₃OD): δ 1.50 (bs, 6H), 1.57 (s,3H), 1.59 (s, 3H), 1.85-2.10 (m, 8H), 2.68 (dd, J=6.5, 14.5, 1H), 2.95(dd, J=4.5, 14.0 Hz, 1H), 3.07 (dd, J=7.0, 13.0 Hz, 1H), 3.17 (dd,J=8.5, 13.5 Hz, 1H), 4.59 (dd, J=4.5, 8.5), 4.97-5.02 (m, 2H), 5.12 (t,J=7.5, 1H), 6.21 (d, J=13.0 Hz, 1H), 6.47 (d, J=13.0 Hz, 1H). ¹³C-NMR(125 MHz, CDCl₃): δ 16.2, 16.3, 17.8, 25.3, 26.0, 27.4, 27.8, 30.3,33.3, 40.8, 40.9, 54.0, 121.5, 125.1, 125.5, 132.1, 133.3, 134.4, 136.3,140.7, 167.7, 168.0, 172.9; ES-MS: mass calcd for Chemical Formula:C₂₂H₃₃NO₅S 423.6. Found (M+Na) m/z 446.

Example 3

Synthesis of(4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2-methylene-4-oxobutanoicacid) (Compound F)

S-trans, trans-Farnesyl-L-cysteine (500 mg, 1.54 mmol) was dissolved inmixture of THF and a first portion of K₂CO₃ (3 mmol) and the resultingsolution was cooled to 5° C. with vigorous stirring. To this stirredsolution was added 3-methylenedihydro-2,5-furandione (302 mg, 3.07 mmol)portionwise while maintaining the pH at 9.0-10.0 with another portion ofK₂CO₃ (3 mmol). The mixture was stirred at room temperature for 3 h.HPLC analysis showed completion of the reaction. The pH of the reactionmixture then adjusted to 2.0 by the addition of 2 N HCl solution. Theacidic solution was extracted three times with 15 mL of ethyl acetate.The combined organic extract was washed with water, brine and dried overNa₂SO₄, the solvent was removed under reduced pressure to afford crudeCompound F, which was further purified by preparative HPLC (552 mg, 82%)to yield Compound F. ¹H-NMR (500 MHz, CDCl₃): δ 1.59 (s, 6H), 1.67 (s,3H), 1.68 (s, 3H), 2.05 (m, 8H), 2.88 (dd, J=6.5, 14.0, 1H), 2.95 (dd,J=6.5, 14.0, 1H), 3.17-3.15 (m, 2H), 3.36 (d, J=14 Hz, 1H), 4.77 (dd,J=6, 12.5 Hz, 1H), 5.09 (bt, 2H), 5.22 (t, J=7.5 Hz, 1H), 5.93 (s, 1H),6.46 (s, 1H). ¹³C-NMR (125 MHz, CDCl₃): δ 16.0, 16.2, 17.7, 25.7, 26.7,29.9, 32.8, 39.6, 39.7, 40.2, 52.0, 119.4, 123.7, 131.3, 132.0, 135.4,140.3, 170.3, 171.5, 176.0; ES-MS: mass calcd for Chemical Formula:C₂₃H₃₅NO₅S 437.6. Found (M+Na) m/z 446.

Example 4

Synthesis of(5-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-5-oxopentanoicacid) (Compound E)

S-trans, trans-Farnesyl-L-cysteine (500 mg, 1.54 mmol) was dissolved inmixture of THF (6 mL) and first portion of K₂CO₃ (3 mmol) and theresulting solution was cooled to 5° C. with vigorous stirring. To thisstirred solution glutaric anhydride (263 mg, 2.30 mmol) was added slowlywhile maintaining the pH at 9.0-9.5 with another portion of K₂CO₃ (3mmol). The mixture was stirred at RT for 3 h, TLC showed completion ofthe reaction. The pH of the reaction mixture then adjusted to 2.0 by theaddition of 2 N hydrochloric acid. The acidic solution was extractedwith ethyl acetate (10 mL×3). The combined organic extract was washedwith water, brine and dried over Na₂SO₄, the solvent was removed onrotary evaporator to afford crude Compound E, which was further purifiedby preparative HPLC (459 mg, 68%) to yield Compound E. ¹H-NMR (500 MHz,CD₃OD): δ 1.60 (s, 6H), 1.70 (s, 3H), 1.72 (s, 3H), 2.02-1.95 (m, 4H),2.15-2.05 (m, 4H), 2.32 (t, 2H), 2.40 (t, 2H), 2.75 (m, 2H), 3.05 (dd,1H), 3.15 (dd, 1H), 3.30 (d, 2H), 4.60 (dd, 1H), 5.14 (t, 2H), 5.25 (t,1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.1, 17.7, 18.4, 22.2, 26.3, 27.4,27.7, 35.7, 40.6, 53.3, 121.5, 125.2, 125.4, 131.8, 136.1, 140.1, 173.7,175.3, 177.6; ES-MS: mass calcd for Chemical Formula: C₂₃H₃₇NO₅S 439.6.Found (M+Na) m/z 462.3.

Example 5

Synthesis of a mixture of(R)-5-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2-(1,3-dioxoisoindolin-2-yl)-5-oxopentanoicacid) (Compound C-1) and(S)-5-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2-(1,3-dioxoisoindolin-2-yl)-5-oxopentanoicacid) (Compound C-2)

S-trans, trans-Farnesyl-L-cysteine (500 mg, 1.54 mmol) was dissolved inmixture of THF (6 mL) and first portion of K₂CO₃ (3 mmol) and theresulting solution was cooled to 5° C. with vigorous stirring. To thisstirred solution was added N-phthaloyl-DL-glutamic anhydride (599 mg,2.31 mmol) as portionwise while maintaining the pH at 9.0-10.0 withanother portion of K₂CO₃ (2 mmol). The mixture was stirred at RT for 3h, TLC/HPLC showed completion of the reaction. The pH of the reactionmixture was adjusted to 2.0 by adding 2 N HCl solution. The acidicsolution was extracted three times with 15 mL of ethyl acetate. Thecombined organic extract was washed with water, brine and dried overNa₂SO₄, and concentrated to afford a crude mixture. The resultingmixture was further purified by preparative HPLC (734 mg, 82%) to yielda mixture of Compound C-1 (the R-R isomer) and Compound C-2 (the S-Risomer), wherein the ratio of C-1 to C-2 is 1:1. ¹H-NMR (500 MHz,CDCl₃): δ 1.50 (s, 3H), 1.52 (s, 3H), 1.55 (s, 1.5H), 1.56 (s, 1.5H),1.60 (s, 3H), 1.86-1.98 (m, 8H), 2.33-2.56 (m, 4H), 2.75 (dd, J=5.0,15.0 Hz, 1H), 2.93 (dd, J=5.0, 15.0 Hz, 1H), 3.03-3.13 (m, 2H), 4.63(dd, J=5.0, 10.0 Hz, 1H), 4.92-5.00 (m, 2H), 5.10 (dd, J=5.0, 15.0 Hz,1H), 6.87 (d, J=5.0 Hz, 0.5H), 6.99 (d, J=5.0 Hz, 0.5H), 7.63-7.65 (m,2H), 7.74-7.77 (m, 2H), 9.30 (broad, 2H). ¹³C-NMR (125 MHz, CDCl₃): δ16.06, 16.17, 16.19, 17.76, 25.20, 25.41, 25.78, 26.48, 26.73, 29.74,29.76, 32.48, 32.63, 32.79, 32.93, 39.66, 39.72, 51.13, 51.19, 51.90,52.11, 119.38, 123.76, 123.81, 124.36, 131.38, 131.60, 131.61, 134.39,134.45, 135.31, 135.34, 140.18, 140.21, 167.78, 167.91, 172.55, 172.72,173.17, 173.35, 174.46, 174.62; ES-MS: mass calcd for Chemical Formula:C₃₁H₄₀N₂O₇S 584.72. Found (M+) m/z 585.3, (M+Na) m/z 607.3.

Example 5a

Alternate Synthesis of a mixture of(R)-5-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2-(1,3-dioxoisoindolin-2-yl)-5-oxopentanoicacid) (Compound C-1) and(S)-5-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2-(1,3-dioxoisoindolin-2-yl)-5-oxopentanoicacid) (Compound C-2)

To a solution of S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) andracemic mixture of N-phthaloyl-glutamic anhydride, i.e.N-phthaloyl-DL-glutamic anhydride (259 mg, 1 mmol) in CH₂Cl₂ (10 mL) wasadded N,N-diisopropyl-ethyl-amine (0.87 mL, 5 mmol). The solution wasstirred at room temperature for 2 h. The reaction was quenched with 1NHCl (10 mL) and the pH was adjusted to 2.0-3.0. The mixtures wereextracted with ethyl acetate (15 mL×3). The organic layer was dried overNa₂SO₄ and concentrated in vacuo and the residue was purified bypreparative HPLC (311 mg, 53%) to yield a mixture of Compound C-1 andCompound C-2, identical to the isomeric mixture obtained in Example 5,wherein the ratio of C-1 to C-2 is 1:1. ¹H-NMR (500 MHz, CDCl₃): δ 1.50(s, 3H), 1.52 (s, 3H), 1.55 (s, 1.5H), 1.56 (s, 1.5H), 1.60 (s, 3H),1.86-1.98 (m, 8H), 2.33-2.56 (m, 4H), 2.75 (dd, J=5.0, 15.0 Hz, 1H),2.93 (dd, J=5.0, 15.0 Hz, 1H), 3.03-3.13 (m, 2H), 4.63 (dd, J=5.0, 10.0Hz, 1H), 4.92-5.00 (m, 2H), 5.10 (dd, J=5.0, 15.0 Hz, 1H), 6.87 (d,J=5.0 Hz, 0.5H), 6.99 (d, J=5.0 Hz, 0.5H), 7.63-7.65 (m, 2H), 7.74-7.77(m, 2H), 9.30 (broad, 2H). ¹³C-NMR (125 MHz, CDCl₃): δ 16.06, 16.17,16.19, 17.76, 25.20, 25.41, 25.78, 26.48, 26.73, 29.74, 29.76, 32.48,32.63, 32.79, 32.93, 39.66, 39.72, 51.13, 51.19, 51.90, 52.11, 119.38,123.76, 123.81, 124.36, 131.38, 131.60, 131.61, 134.39, 134.45, 135.31,135.34, 140.18, 140.21, 167.78, 167.91, 172.55, 172.72, 173.17, 173.35,174.46, 174.62; ES-MS: mass calcd for Chemical Formula: C₃₁H₄₀N₂O₇S584.72. Found (M+) m/z 585.3, (M+Na) m/z 607.3.

Example 5b

Synthesis of((S)-5-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2-(1,3-dioxoisoindolin-2-yl)-5-oxopentanoicacid) (Compound C-2)

To a solution of S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) andN-phthaloyl-L-glutamic anhydride (259 mg, 1 mmol) in CH₂Cl₂ (10 mL) wasadded N,N-diisopropyl-ethyl-amine (0.87 mL, 5 mmol). The solution wasstirred at room temperature for 2 h. The reaction was quenched with 1NHCl (10 mL) and the pH was adjusted to 2.0-3.0. The mixtures wereextracted with ethyl acetate (15 mL×3). The organic layer was dried overNa₂SO₄ and concentrated in vacuo and the residue was purified bypreparative HPLC (350 mg, 60%) to yield Compound C-2, which is identicalto the S-R stereoisomer of the Compound C racemate in Examples 5 and 5a.¹H-NMR (500 MHz, CDCl₃): δ 1.50 (s, 3H), 1.52 (s, 3H), 1.55 (s, 3H),1.60 (s, 3H), 1.86-1.98 (m, 8H), 2.33-2.43 (m, 2H), 2.54-2.57 (m, 2H),2.78 (dd, J=5.0, 15.0 Hz, 1H), 2.91 (dd, J=5.0, 15.0 Hz, 1H), 3.06 (dd,J=5.0, 10.0 Hz, 1H), 3.14 (dd, J=5.0, 10.0 Hz, 1H), 4.65 (dd, J=5.0,10.0 Hz, 1H), 4.96 (t, J=5.0 Hz, 1H), 5.00 (m, 2H), 5.11 (t, J=5.0 Hz,1H), 6.79 (d, J=10.0 Hz, 1H), 7.63-7.65 (m, 2H), 7.74-7.76 (m, 2H), 8.00(broad, 2H). ¹³C-NMR (125 MHz, CDCl₃): δ 16.06, 16.18, 17.76, 25.19,25.78, 26.47, 26.73, 29.77, 32.59, 32.79, 39.65, 39.72, 51.10, 51.85,119.37, 123.76, 123.80, 124.36, 131.39, 131.62, 134.40, 135.36, 140.24,167.75, 172.78, 173.05, 174.51; ES-MS: mass calcd for Chemical Formula:C₃₁H₄₀N₂O₇S 584.72. Found (M+) m/z 585.3, (M+Na) m/z 607.3.

Example 6

Synthesis of((R,14E,18E)-15,19,23-trimethyl-4,8-dioxo-3-oxa-12-thia-7,9-diazatetracosa-14,18,22-triene-10-carboxylicacid) (Compound D)

S-trans, trans-Farnesyl-L-cysteine (325 mg, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). Ethyl-3-isocyanato-propionate (143, 1 mmol) was added to thereaction mixture. The reaction solution was stirred at room temperatureovernight and the solvent was removed by rotary evaporation. Theremaining residue was dissolved in ethyl acetate (100 mL) and washedwith 1 N HCl solution (50 mL×2). The ethyl acetate solution was driedover Na₂SO₄ and concentrated to a crude reaction mixture. The resultingmixture was purified by HPLC (200 mg, 43%) to yield Compound D. ¹H-NMR(500 MHz, MeOH-d₄): δ 1.28 (t, J=7.5 Hz, 3H), 1.62 (s, 3H), 1.63 (s,3H), 1.69 (s, 3H), 1.70 (s, 3H), 1.98 (m, 2H), 2.06 (m, 6H), 2.52 (t,J=6.5, 2H), 2.79 (dd, J=7.0, 14.0 Hz, 1H), 2.93 (dd, J=4.5, 13.5 Hz,1H), 3.17 (dd, J=7.5, 13.5 Hz, 1H), 3.28 (dd, J=8.5, 13.5 Hz, 1H), 3.41(t, J=6.5 Hz, 3H), 4.16 (q, J=7.5 Hz, 2H), 4.50 (dd, J=4.5, 6.5 Hz, 1H),5.11 (m, 2H), 5.23 (t, J=7.5, 1H). ¹³C-NMR (125 MHz, MeOH-d₄): δ 14.6,16.1, 16.2, 17.8, 26.0, 27.4, 27.8, 30.5, 34.4, 35.9, 36.7, 40.8, 40.9,54.0, 61.7, 121.7, 125.1, 125.5, 132.1, 136.3, 140.5, 160.2, 173.8,175.0; ES-MS: mass calcd for Chemical Formula: C₂₄H₄₀N₂O₅S 468.7. Found(M+Na) m/z 491.3.

Example 7

Synthesis of((R)-2-(3-(2-carboxyethyl)ureido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound G)

In a 100 mL round bottom flask, Compound D of Example 6 (100 mg, 0.21mmol) was dissolved in THF (10 mL). LiOH (500 mg, 20 mmol) in water (5mL) was added to the reaction solution. The reaction mixture was stirredat room temperature overnight. Ethyl acetate (100 mL) was added to thereaction mixture. The reaction mixture was acidified by 1 N HCl solution(pH=4.0). The organic portion was separated and dried over Na₂SO₄,concentrated and purified by HPLC (40 mg, 41%) to yield Compound G.¹H-NMR (500 MHz, MeOH-d₄): δ 1.62 (bs, 6H), 1.69 (s, 3H), 1.70 (s, 3H),1.99 (m, 2H), 2.08 (m, 6H), 2.51 (t, J=6.5, 2H), 2.79 (dd, J=7.0, 14.0Hz, 1H), 2.93 (dd, J=4.5, 8.1 Hz, 1H), 3.17 (dd, J=7.0, 13.0 Hz, 1H),3.28 (dd, J=9.0, 15.0 Hz, 1H), 3.40 (t, J=6.5 Hz, 3H), 4.50 (dd, J=5.0,6.5 Hz, 1H), 5.11 (m, 2H), 5.23 (t, J=7.5, 1H). ¹³C-NMR (125 MHz,MeOH-d₄): δ 16.1, 16.2, 17.8, 26.0, 27.4, 27.8, 30.5, 34.4, 35.7, 36.8,40.8, 40.9, 54.1, 121.7, 125.2, 125.5, 132.1, 136.3, 140.5, 160.3,175.1, 175.7; ES-MS: mass calcd for Chemical Formula: C₂₂H₃₆N₂O₅S 440.6.Found (M+Na) m/z 463.3.

Example 8

Synthesis of Compound I as a mixture of((1R,2S)-2-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylcarbamoyl)cyclopropanecarboxylicacid) (Compound I-1) and((1S,2R)-2-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylcarbamoyl)cyclopropanecarboxylicacid) (Compound I-2)

In a 100 mL round bottom flask, S-trans, trans-farnesyl-L-cysteine (325mg, 1 mmol) and N,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixedin CH₂Cl₂ (10 mL). 3-oxabicyclo[3.1.0]hexane-2,4-dione (112 mg, 1.0mmol) was added to the reaction mixture. The reaction solution wasstirred at room temperature overnight and the solvent was removed byrotary evaporation. The remaining residue was dissolved in ethyl acetate(100 mL) and washed with 1 N HCl solution (10 mL). The ethyl acetatesolution was dried over Na₂SO₄ to afford a concentrated crude mixture.The crude mixture was purified by preparative HPLC (250 mg, 57%) toyield mixture of Compound I-1 and Compound I-2, wherein the ratio of I-1to I-2 is 1:1. ¹H-NMR (500 MHz, CD₃OD): δ 1.32 (m, 1H), 1.56 (m, 1H),1.62 (bs, 6H), 1.69 (s, 3H), 1.71 (s, 3H), 1.97 (t, J=7.0 Hz, 2H),2.04-2.22 (m, 8H), 2.73-2.78 (m, 1H), 2.95-3.00 (m, 1H), 3.13-3.18 (m,1H), 3.25-3.33 (m, 1H), 4.60 (m, 1H), 5.09 (m, 2H), 5.19 (t, J=7.5 Hz,1H). ¹³C-NMR (125 MHz, CDCl₃): δ 12.5, 12.6, 16.2, 16.3, 17.8, 22.6,22.8, 24.1, 24.2, 26.0, 27.4, 27.8, 30.2, 33.4, 33.5, 40.8, 40.9, 53.6,121.6, 125.1, 125.5, 132.1, 136.2, 140.5, 172.3, 173.7, 173.9, 174.5;ES-MS: mass calcd for Chemical Formula: C₂₃H₃₅NO₅S 437.6. Found (M+Na)m/z 460.3.

Example 9

Synthesis of((6S,9R,13E,17E)-6-(hydroxymethyl)-2,2,14,18,22-pentamethyl-4,7-dioxo-3-oxa-11-thia-5,8-diazatricosa-13,17,21-triene-9-carboxylicacid) (Compound N-55)

In a 100 mL round bottom flask, N-Boc-L-serine (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture. The reaction mixture was stirred at roomtemperature overnight and the CH₂Cl₂ was removed by rotary evaporation.The remaining residue was dissolved in ethyl acetate (50 mL). Theresulting organic solution was washed with an NH₄Cl saturated solution(50 mL), dried over Na₂SO₄, and concentrated to afford a crude mixture.The crude mixture was purified by preparative HPLC (51 mg, 10%) to yieldCompound N-55. ¹H-NMR (500 MHz, CD₃OD): δ 1.47 (s, 9H), 1.62 (bs, 6H),1.69 (s, 3H), 1.71 (s, 3H), 1.99 (t, J=7 Hz, 2H), 2.05-2.22 (m, 6H),2.81 (dd, J=7.5, 14 Hz, 1H), 3.01 (dd, J=4, 14.5 Hz, 1H), 3.17 (dd, J=7,13 Hz, 1H), 3.27 (dd, J=8, 12.5 Hz, 1H), 3.33 (bs, 2H), 3.74-3.80 (m,2H), 4.22 (t, J=5 Hz, 1H), 4.63 (dd, J=5, 7.5 Hz, 1H), 5.11 (m, 2H),5.23 (t, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz, CDCl₃): δ 16.2, 16.3, 17.8,26.0, 27.4, 27.8, 28.7, 30.4, 33.6, 40.8, 40.9, 53.3, 58.0, 59.6, 63.4,80.9, 121.6, 125.2, 125.5, 132.1, 136.3, 140.6, 157.8, 173.0, 173.8;ES-MS: mass calcd for Chemical Formula: C₂₆H₄₄N₂O₆S 512.7. Found (M+Na)m/z 535.4.

Example 10

Synthesis of((6S,9R,13E,17E)-6-((S)-1-hydroxyethyl)-2,2,14,18,22-pentamethyl-4,7-dioxo-3-oxa-11-thia-5,8-diazatricosa-13,17,21-triene-9-carboxylicacid) (Compound N-56)

In a 100 mL round bottom flask, N-Boc-L-threonine (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (145 mg, 28%) to yield Compound N-56. ¹H-NMR (500 MHz, CD₃OD): δ1.21 (d, J=6 Hz, 3H), 1.48 (s, 9H), 1.62 (bs, 6H), 1.69 (s, 3H), 1.71(s, 3H), 1.99 (t, J=7.5 Hz, 2H), 2.05-2.22 (m, 6H), 2.82 (dd, J=7.5, 14Hz), 3.01 (dd, J=5, 13.5 Hz, 1H), 3.17 (dd, J=7, 13.5 Hz, 1H), 3.29 (dd,J=8.5, 13.5 Hz, 1H), 3.33 (bs, 2H), 4.09-4.16 (m, 2H), 4.65 (dd, J=5, 7Hz, 1H), 5.11 (m, 2H), 5.23 (t, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz, CDCl₃):δ 16.2, 16.3, 17.8, 19.9, 26.0, 27.4, 27.8, 28.6, 30.4, 33.5, 40.8,40.9, 53.4, 61.3, 63.0, 68.7, 80.8, 121.6, 125.15, 125.5, 132.12,136.26, 140.6, 157.9, 173.2, 173.6; ES-MS: mass calcd for ChemicalFormula: C₂₇H₄₆N₂O₆S 526.7. Found (M+Na) m/z 549.4.

Example 11

Synthesis of((6S,9R,13E,17E)-2,2,14,18,22-pentamethyl-6-(2-(methylthio)ethyl)-4,7-dioxo-3-oxa-11-thia-5,8-diazatricosa-13,17,21-triene-9-carboxylicacid) (Compound N-57)

In a 100 mL round bottom flask, N-Boc-L-methionine (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-Farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture. The reaction mixture was stirred at roomtemperature overnight. CH₂Cl₂ was removed by rotary evaporation. Theresulting residue was dissolved in ethyl acetate (50 mL). The organicsolution was washed with an NH₄Cl saturated solution (50 mL), dried overNa₂SO₄, and concentrated to afford a crude mixture. The crude mixturewas purified by preparative HPLC (290 mg, 52%) to yield Compound N-57.¹H-NMR (500 MHz, CDCl₃): δ 1.47 (s, 9H), 1.62 (bs, 6H), 1.69 (s, 3H),1.72 (s, 3H), 1.90-1.93 (m, 1H), 1.99 (t, J=7.5 Hz, 2H), 2.03-2.16 (m,8H), 2.50-2.68 (m, 2H), 2.80 (dd, J=8, 14 Hz, 1H), 3.02 (dd, J=4.5, 14Hz, 1H), 3.18 (dd, J=7, 13 Hz, 1H), 3.27 (dd, J=8, 13 Hz, 1H), 3.25 (dd,J=5, 8 Hz, 1H), 4.60 (dd, J=4.5, 8, 1H), 5.10-5.15 (m, 2H), 5.24 (t,J=7.5, 1H). ¹³C-NMR (125 MHz, CDCl₃): δ 15.3, 16.2, 16.3, 17.8, 26.0,27.4, 27.8, 28.8, 30.3, 31.0, 33.0, 33.4, 40.8, 40.9, 53.3, 55.1, 80.8,121.6, 125.2, 125.5, 132.1, 136.3, 140.6, 157.8, 173.6, 174.6; ES-MS:mass calcd for Chemical Formula: C₂₈H₄₈N₂O₅S₂ 556.3. Found (M+Na) m/z279.2.

Example 12

Synthesis of((6S,9R,13E,17E)-6-isobutyl-2,2,14,18,22-pentamethyl-4,7-dioxo-3-oxa-11-thia-5,8-diazatricosa-13,17,21-triene-9-carboxylicacid) (Compound N-58)

In a 100 mL round bottom flask, N-Boc-L-leucine (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture. The reaction mixture was stirred at roomtemperature overnight, the CH₂Cl₂ was removed by rotary evaporation. Theresulting residue was dissolved in ethyl acetate (50 mL). The organicsolution was washed with an NH₄Cl saturated solution (50 mL), dried overNa₂SO₄, and concentrated to afford a crude mixture. The crude mixturewas purified by preparative HPLC (200 mg, 37%) to yield Compound N-58.¹H-NMR (500 MHz, CDCl₃): δ 0.95 (d, J=6.5 Hz, 3H), 0.98 (d, J=6.5 Hz,3H), 1.47 (s, 9H), 1.62 (bs, 6H), 1.51-1.59 (m, 1H), 1.69 (s, 3H), 1.72(s, 3H), 1.99 (t, J=7.5 Hz, 2H), 2.05-2.14 (m, 8H), 2.79 (dd, J=7.5,14.2, 1H), 3.00 (dd, J=4.8, 13.9 Hz, 1H), 3.17 (dd, J=7.6, 13.0 Hz, 1H),3.25 (dd, J=8.2, 12.9 Hz, 1H), 4.15 (dd, J=5.4, 9.8 Hz, 1H), 4.60 (dd,J=4.9, 8.0 Hz, 1H), 5.09-5.25 (m, 2H), 5.23 (t, J=7.6 Hz, 1H). ¹³C-NMR(125 MHz, CDCl₃): δ 16.2, 16.3, 17.8, 22.0, 23.5, 25.9, 27.4, 27.8,28.8, 30.4, 33.6, 40.8, 40.9, 42.3, 53.3, 54.6, 80.6, 121.6, 125.2,125.5, 132.1, 136.2, 140.6, 157.8, 173.6, 175.6; ES-MS: mass calcd forChemical Formula: C₂₉H₅₀N₂O₅S 538.3. Found (M+Na) m/z 561.4.

Example 13

Synthesis of((6S,9R,13E,17E)-6-(R)-sec-butyl-2,2,14,18,22-pentamethyl-4,7-dioxo-3-oxa-11-thia-5,8-diazatricosa-13,17,21-triene-9-carboxylicacid) (Compound N-59)

In a 100 mL round bottom flask, N-Boc-L-isoleucine (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and was stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (210 mg, 39%) to yield Compound N-59. ¹H-NMR (500 MHz, CDCl₃): δ0.92 (t, J=7.4 Hz, 3H), 0.97 (d, J=6.9 Hz, 3H), 1.19 (m, 1H), 1.47 (s,9H), 1.61 (m, 1H), 1.62 (bs, 6H), 1.69 (s, 3H), 1.72 (s, 3H), 1.82 (m,1H), 1.99 (t, J=7.5 Hz, 2H), 2.05-2.14 (m, 8H), 2.78 (dd, J=8.0, 14.0,1H), 3.01 (dd, J=4.9, 14.0 Hz, 1H), 3.18 (dd, J=7.4, 13.1 Hz, 1H), 3.27(dd, J=8.4, 13.1 Hz, 1H), 4.00 (d, J=7.3 Hz, 1H), 4.62 (dd, J=4.9, 8.0Hz, 1H), 5.10-5.17 (m, 2H), 5.24 (t, J=7.4 Hz, 1H). ¹³C-NMR (125 MHz,CDCl₃): δ 11.6, 16.0, 16.2, 16.3, 17.9, 25.8, 26.0, 27.5, 27.8, 28.8,30.4, 33.6, 38.5, 40.8, 40.9, 53.4, 60.7, 80.6, 121.6, 125.2, 125.5,132.1, 136.3, 140.5, 157.9, 173.6, 174.4; ES-MS: mass calcd for ChemicalFormula: C₂₉H₅₀N₂O₅S 538.3. Found (M+Na) m/z 561.4.

Example 14

Synthesis of((R)-2-((S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-8)

In a 100 mL round bottom flask, N-Boc-L-proline (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture. The reaction mixture was stirred at roomtemperature overnight. CH₂Cl₂ was removed by rotary evaporation. Theresulting residue was dissolved in ethyl acetate (50 mL). The organicsolution was washed with an NH₄Cl saturated solution (50 mL), dried overNa₂SO₄, and concentrated to afford a crude mixture. The crude mixturewas purified by preparative HPLC (232 mg, 44%) to yield Compound N-8.¹H-NMR (500 MHz, CDCl₃): δ 1.46 (s, 9H), 1.62 (bs, 6H), 1.69 (s, 3H),1.72 (s, 3H), 1.89 (bs, 1H), 1.98 (t, J=7.5 Hz, 2H), 2.05-2.14 (m, 6H),2.24-2.26 (m, 1H), 2.79 (dd, J=8.0, 14.0, 1H), 3.03 (bd, J=13.0 Hz, 1H),3.15 (dd, J=7.5, 13.0 Hz, 1H), 3.27 (dd, J=8.0, 13.0 Hz, 1H), 3.40 (m,1H), 3.66 (bs, 1H), 4.28 (bs, 1H), 4.60 (dd, J=4.9, 8.0 Hz, 1H),5.11-5.14 (m, 2H), 5.24 (t, J=7.4 Hz, 1H). ¹³C-NMR (125 MHz, CDCl₃): δ16.2, 16.3, 17.8, 24.2, 24.5, 25.3, 25.9, 27.4, 27.8, 28.7, 30.1, 30.4,30.7, 31.1, 32.5, 33.5, 40.8, 40.9, 47.9, 48.3, 53.1, 53.4, 61.4, 61.9,81.4, 81.7, 121.5, 125.1, 125.5, 132.1, 136.3, 140.5, 156.2, 173.6,175.8; ES-MS: mass calcd for Chemical Formula: C₂₈H₄₆N₂O₅S 522.3. Found(M+Na) m/z 545.3.

Example 15

Synthesis of((6S,9R,13E,17E)-2,2,6,14,18,22-hexamethyl-4,7-dioxo-3-oxa-11-thia-5,8-diazatricosa-13,17,21-triene-9-carboxylicacid) (Compound N-3)

In a 100 mL round bottom flask, N-Boc-L-alanine (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (170 mg, 35%) to yield Compound N-3. ¹H-NMR (500 MHz, CDCl₃): δ1.37 (bt, 3H), 1.44 (s, 9H), 1.60 (bs, 6H), 1.68 (bs, 6H), 1.99 (t,J=8.2 Hz, 2H), 2.04-2.10 (m, 6H), 2.89 (dd, J=6.3, 13.9 Hz, 1H), 3.01(dd, J=7.5, 14.1 Hz, 1H), 3.14-3.24 (m, 2H), 4.31-4.39 (m, 1H),4.74-4.77 (m, 1H), 5.08-5.10 (m, 2H), 5.23 (t, J=7.5 Hz, 1H). ¹³C-NMR(125 MHz, CDCl₃): δ 16.1, 16.2, 17.8, 18.6, 19.1, 25.7, 26.5, 26.7,28.5, 30.0, 33.0, 33.1, 39.7, 49.7, 49.8, 51.7, 51.8, 51.9, 76.8, 77.0,77.3, 80.5, 80.7, 119.6, 123.8, 124.3, 131.3, 135.3, 140.1, 155.7,155.9, 173.0, 173.1; ES-MS: mass calcd for Chemical Formula: C₂₆H₄₄N₂O₅S496.7. Found (M+Na) m/z 519.4.

Example 16

Synthesis of((R,13E,17E)-2,2,14,18,22-pentamethyl-4,7-dioxo-3-oxa-11-thia-5,8-diazatricosa-13,17,21-triene-9-carboxylicacid) (Compound N-4)

In a 100 mL round bottom flask, N-Boc-L-glycine (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes, S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedand the reaction mixture was stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (78 mg, 18%) to yield Compound N-4. ¹H-NMR (500 MHz, CDCl₃): δ 1.45(s, 9H), 1.60 (bs, 6H), 1.67 (s, 3H), 1.68 (s, 3H), 1.97 (t, J=7.5 Hz,2H), 2.01-2.15 (m, 6H), 2.85 (bd, J=13 Hz, 1H), 3.01 (bd, J=13 Hz, 1H),3.14-3.23 (m, 2H), 3.81 (d, J=16.5 Hz), 4.01 (d, J=12 Hz, 1H), 4.77 (m,1H), 5.09 (m, 2H), 5.30 (1H). ¹³C-NMR (125 MHz, CDCl₃): δ 16.0, 16.1,17.7, 25.7, 26.5, 26.7, 28.4, 30.0, 32.9, 39.7, 39.7, 43.8, 51.7, 80.7,119.5, 123.8, 124.3, 131.4, 135.4, 140.1, 156.3, 169.8, 172.9; ES-MS:mass calcd for Chemical Formula: C₂₅H₄₂N₂O₅S 482.3. Found (M+Na) m/z505.1.

Example 17

Synthesis of((6S,9R,13E,17E)-2,2,14,18,22-pentamethyl-4,7-dioxo-6-phenyl-3-oxa-11-thia-5,8-diazatricosa-13,17,21-triene-9-carboxylicacid) (Compound N-5)

In a 100 mL round bottom flask, N-Boc-L-phenylglycine (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30 minutesand S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was added. Thereaction mixture was stirred at room temperature overnight. CH₂Cl₂ wasremoved by rotary evaporation. The resulting residue was dissolved inethyl acetate (50 mL). The organic solution was washed with an NH₄Clsaturated solution (50 mL), dried over Na₂SO₄, and concentrated toafford a crude mixture. The crude mixture was purified by preparativeHPLC (118 mg, 21%) to yield Compound N-5. ¹H-NMR (500 MHz, CD₃OD): δ1.47 (s, 9H), 1.62 (s, 6H), 1.69 (s, 6H), 1.99 (t, J=7.0 Hz, 2H),2.03-2.13 (m, 6H), 2.81 (dd, J=8.1, 13.9 Hz, 1H), 3.01 (dd, J=6.8, 12.1Hz, 1H), 3.16 (dd, J=7.6, 13.2 Hz, 1H), 3.26 (dd, J=8.5, 13.4 Hz), 4.62(bt, J=5.5), 5.09-5.12 (m, 2H), 5.22 (t, J=7.9 Hz, 1H), 7.29-7.37 (m,3H), 7.46 (d, J=7.3 Hz, 2H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.2, 16.3,25.9, 27.3, 27.8, 28.7, 30.3, 33.4, 33.5, 40.8, 40.9, 53.6, 59.9, 81.1,121.6, 125.2, 125.5, 128.6, 129.1, 129.7, 132.1, 136.3, 139.1, 140.5,158.3, 173.0, 173.4; ES-MS: mass calcd for Chemical Formula: C₃₁H₄₆N₂O₅S558.8. Found (M+Na) m/z 581.4.

Example 18

Synthesis of((R,14E,18E)-2,2,15,19,23-pentamethyl-4,8-dioxo-3-oxa-12-thia-5,9-diazatetracosa-14,18,22-triene-10-carboxylicacid) (Compound N-6)

In a 100 mL round bottom flask, N-Boc-L-beta-alanine (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedand stirred at room temperature overnight. CH₂Cl₂ was removed by rotaryevaporation. The resulting residue was dissolved in ethyl acetate (50mL). The organic solution was washed with an NH₄Cl saturated solution(50 mL), dried over Na₂SO₄, and concentrated to afford a crude mixture.The crude mixture was purified by preparative HPLC (171 mg, 35%) toyield Compound N-6. ¹H-NMR (500 MHz, CD₃OD): δ 1.45 (s, 9H), 1.60 (s,3H), 1.61 (s, 3H), 1.69 (s, 3H), 1.72 (s, 3H), 1.99 (t, J=7.6 Hz, 2H),2.01-2.15 (m, 6H), 2.47 (t, J=6.8 Hz, 2H), 2.73 (dd, J=8.8, 13.9 Hz,1H), 3.01 (dd, J=4.6, 14.0 Hz, 1H), 3.16 (dd, J=7.3, 13.2 Hz, 1H), 3.28(dd, J=8.4, 13.4 Hz, 1H), 3.33 (m, 2H), 4.61 (dd, J=4.6, 9.0 Hz, 1H),5.10-5.22 (m, 2H), 5.24 (t, J=7.7 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ16.2, 16.3, 17.8, 25.9, 27.4, 27.8, 28.8, 30.2, 33.5, 37.0, 38.0, 40.8,40.9, 53.3, 80.2, 121.6, 125.1, 125.5, 132.1, 136.3, 140.6, 158.3,174.0; ES-MS: mass calcd for Chemical Formula: C₂₆H₄₄N₂O₅S 496.7. Found(M+Na) m/z 519.3.

Example 19

Synthesis of((R,15E,19E)-2,2,16,20,24-pentamethyl-4,9-dioxo-3-oxa-13-thia-5,10-diazapentacosa-15,19,23-triene-11-carboxylicacid) (Compound N-7)

In a 100 mL round bottom flask, N-Boc-L-aminobutanoic acid (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedand stirred at room temperature overnight. CH₂Cl₂ was removed by rotaryevaporation. The resulting residue was dissolved in ethyl acetate (50mL). The organic solution was washed with an NH₄Cl saturated solution(50 mL), dried over Na₂SO₄, and concentrated to afford a crude mixture.The crude mixture was purified by preparative HPLC (142 mg, 32%) toyield Compound N-7. ¹H-NMR (500 MHz, CD₃OD): δ 1.46 (s, 9H), 1.62 (s,3H), 1.63 (s, 3H), 1.69 (s, 3H), 1.72 (s, 3H), 1.77-1.80 (m, 2H), 1.99(t, J=7.5 Hz, 2H), 2.01-2.15 (m, 6H), 2.31 (t, J=7.5 Hz, 2H), 2.73 (dd,J=9.0, 13.9 Hz, 1H), 3.02 (dd, J=4.5, 14.0 Hz, 1H), 3.11 (t, J=6.8 Hz,2H), 3.17 (dd, J=7.3, 13.2 Hz, 1H), 3.29 (dd, J=8.4, 13.2 Hz, 1H), 4.60(dd, J=4.6, 9.0 Hz, 1H), 5.10-5.22 (m, 2H), 5.24 (t, J=7.8 Hz, 1H).¹³C-NMR (125 MHz, CD₃OD): δ 16.2, 16.3, 17.8, 25.9, 27.3, 27.4, 27.8,28.8, 30.2, 33.5, 34.1, 40.7, 40.8, 40.9, 53.3, 80.0, 121.6, 125.1,125.5, 132.1, 136.3, 140.5, 158.6, 174.0, 175.7; ES-MS: mass calcd forChemical Formula: C₂₇H₄₆N₂O₅S 410.3. Found (M+Na) m/z 533.3.

Example 20

Synthesis of((6S,9R,13E,17E)-6-isopropyl-2,2,14,18,22-pentamethyl-4,7-dioxo-3-oxa-11-thia-5,8-diazatricosa-13,17,21-triene-9-carboxylicacid) (Compound N-9)

In a 100 mL round bottom flask, N-Boc-L-valine (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (276 mg, 53%) to yield Compound N-9. ¹H-NMR (500 MHz, CD₃OD): δ0.95 (d, J=6.5 Hz, 3H), 0.99 (d, J=6.5 Hz, 3H), 1.21 (m, 1H), 1.47 (s,9H), 1.62 (bs, 6H), 1.69 (s, 3H), 1.71 (s, 3H), 1.99 (t, J=7.5 Hz, 2H),2.05-2.15 (m, 8H), 2.78 (dd, J=8.5, 14.0, 1H), 3.00 (dd, J=4.5, 14.0 Hz,1H), 3.16 (dd, J=7.5, 13.2 Hz, 1H), 3.27 (dd, J=8.4, 13.2 Hz, 1H), 3.95(d, J=6.5 Hz, 1H), 4.61 (dd, J=4.8, 8.0 Hz, 1H), 5.10-5.14 (m, 2H), 5.51(t, J=7.4 Hz, 1H). ¹³C-NMR (125 MHz, CDCl₃): δ 16.2, 16.3, 17.8, 18.4,19.8, 25.9, 27.4, 27.8, 28.8, 30.3, 32.2, 33.5, 40.8, 40.9, 53.3, 61.5,80.6, 121.6, 125.2, 125.5, 132.1, 136.3, 140.5, 157.9, 173.6, 174.4;ES-MS: mass calcd for Chemical Formula: C₂₈H₄₈N₂O₅S 524.7. Found (M+Na)m/z 547.4.

Example 21

Synthesis of((6R,9R,13E,17E)-6-benzhydryl-2,2,14,18,22-pentamethyl-4,7-dioxo-3-oxa-11-thia-5,8-diazatricosa-13,17,21-triene-9-carboxylicacid) (Compound N-12)

In a 100 mL round bottom flask, N-Boc-D-diphenyl-alanine (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (306 mg, 68%) to yield Compound N-12. ¹H-NMR (500 MHz, CD₃OD): δ1.33 (s, 9H), 1.62 (s, 3H), 1.63 (s, 3H), 1.65 (s, 3H), 1.69 (s, 3H),1.99 (t, J=7.0 Hz, 2H), 2.03-2.16 (m, 8H), 2.39 (dd, J=6.5, 14.0 Hz,1H), 2.52 (dd, J=6.5, 14.0 Hz, 1H), 2.96-3.05 (m, 2H), 4.28-4.36 (m,1H), 4.37 (s, 1H), 5.01 (d, J=11.0 Hz, 1H), 5.10-5.20 (m, 2H), 7.15-7.38(m, 10H). ¹³C-NMR (125 MHz, CDCl₃): δ 16.2, 16.3, 26.0, 27.4, 27.8,28.6, 30.3, 33.2, 40.7, 40.9, 53.3, 53.4, 55.1, 58.6, 80.6, 121.5,125.2, 125.5, 127.7, 127.9, 129.4, 129.6, 129.7, 132.1, 136.3, 140.3,142.4, 142.5, 157.4, 173.1, 173.5; ES-MS: mass calcd for ChemicalFormula: C₃₈H₅₂N₂O₅S 648.4. Found (M+Na) m/z 671.2.

Example 22

Synthesis of((R)-2-(1-(tert-butoxycarbonylamino)cyclopropanecarboxamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-49)

In a 100 mL round bottom flask, N-Boc-amino-cyclopropionic acid (1.0mmol), coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (158 mg, 31%) to yield Compound N-49. ¹H-NMR (500 MHz, CD₃OD): δ1.05-1.08 (m, 2H), 1.41-1.46 (m, 2H), 1.49 (s, 9H), 1.62 (s, 3H), 1.63(s, 3H), 1.69 (s, 3H), 1.70 (s, 3H), 1.99 (t, J=8.2 Hz, 2H), 2.05-2.16(m, 6H), 2.92 (dd, J=5.5, 13.5 Hz, 1H), 3.01 (dd, J=7.5, 14.0 Hz, 1H),3.16 (dd, J=7.5, 13.0 Hz, 1H), 3.27 (dd, J=8.5, 13.0 Hz, 1H), 4.61 (bs,1H), 5.12 (dd, J=7.5, 15.5 Hz, 2H), 5.23 (t, J=7.5 Hz, 1H). ¹³C-NMR (125MHz, CDCl₃): δ 16.1, 16.3, 17.8, 25.9, 27.4, 27.8, 30.7, 33.7, 40.8,40.9, 53.7, 53.8, 121.6, 125.1, 125.5, 132.1, 136.3, 140.6, 158.1,173.6; ES-MS: mass calcd for Chemical Formula: C₂₇H₄₄N₂O₅S 508.7. Found(M+Na) m/z 531.4.

Example 23

Synthesis of((6S,9R,13E,17E)-6-cyclohexyl-2,2,14,18,22-pentamethyl-4,7-dioxo-3-oxa-11-thia-5,8-diazatricosa-13,17,21-triene-9-carboxylicacid ((Compound N-60)

In a 100 mL round bottom flask, N-Boc-L-cyclohexyl-Glycine (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (260 mg, 58%) to yield Compound N-60. ¹H-NMR (500 MHz, CD₃OD): δ1.05-1.30 (m, 6H), 1.47 (s, 9H), 1.61-1.77 (m, 5H), 1.62 (bs, 6H), 1.69(s, 3H), 1.76 (s, 3H), 1.99 (t, J=7.5 Hz, 2H), 2.06-2.17 (m, 8H), 2.78(dd, J=8.0, 14.0, 1H), 3.01 (dd, J=5.0, 14.0 Hz, 1H), 3.18 (dd, J=7.5,13.0 Hz, 1H), 3.27 (dd, J=8.0, 13.0 Hz, 1H), 3.96 (d, J=6.5 Hz, 1H),4.61 (dd, J=5.0, 8.0 Hz, 1H), 5.12 (dd, J=8.0, 17.0 Hz, 2H), 5.24 (t,J=7.5 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.2, 16.3, 17.8, 26.0, 27.1,27.2, 27.3, 27.5, 27.8, 28.8, 33.5, 40.8, 40.9, 41.8, 53.3, 61.0, 62.5,80.6, 121.6, 125.2, 125.5, 132.1, 136.3, 140.5, 157.9, 173.6, 174.3;ES-MS: mass calcd for Chemical Formula: C₃₁H₅₂N₂O₅S 564.8. Found (M+Na)m/z 587.4.

Example 24

Synthesis of((R)-2-((R)-2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinoline-3-carboxamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-50)

In a 100 mL round bottom flask,N-Boc-(R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (250 mg, 48%) to yield Compound N-50. ¹H-NMR (500 MHz, CD₃OD): δ1.33 (s, 9H), 1.61 (bs, 6H), 1.68 (bs, 6H), 1.69-1.73 (m, 2H), 1.99 (t,J=7.5 Hz, 2H), 2.06-2.17 (m, 8H), 2.73 (dd, J=7.5, 14.0, 1H), 2.87 (dd,J=5.0, 14.0 Hz, 1H), 3.16 (dd, J=7.5, 13.0 Hz, 1H), 3.25 (dd, J=8.0,13.0 Hz, 1H), 3.25-3.30 (m, 2H), 3.73 (dd, J=5.5, 14.0 Hz, 1H),4.55-4.61 (m, 2H), 5.12 (dd, J=8.0, 17.0 Hz, 2H), 5.20 (t, J=7.5 Hz,1H), 7.39-7.88 (m, 3H), 8.20-8.24 (m, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ16.2, 16.3, 17.9, 26.0, 27.4, 27.8, 28.1, 28.7, 30.4, 30.5, 30.9, 33.4,36.6, 37.9, 40.8, 40.9, 53.3, 56.9, 57.7, 121.5, 124.8, 125.1, 125.5,126.5, 126.6, 127.3, 128.6, 128.7, 128.8, 129.3, 129.9, 132.1, 133.5,134.7, 135.5, 136.3, 140.6, 157.5, 173.7, 174.2; ES-MS: mass calcd forChemical Formula: C₃₃H₄₈N₂O₅S 584.8. Found (M+Na) m/z 607.4.

Example 25

Synthesis of a mixture of((R)-2-((S)-2-acetamido-3-hydroxypropanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) and((R)-2-((R)-2-acetamido-3-hydroxypropanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid)) (Compound N-23)

In a 100 mL round bottom flask, N-acetyl-L-serine (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (160 mg, 35%) to yield a 1:1 ratio mixture of R-R and S-R isomersof Compound N-23, similar to Compound C in Examples 5 and 5a. ¹H-NMR(500 MHz, CD₃OD): δ 1.50 (s, 6H), 1.57 (s, 3H), 1.58 (s, 3H), 1.85-2.03(m, 11H), 2.72-2.79 (m, 1H), 2.87-2.96 (m, 1H), 3.07-3.14 (m, 2H),3.65-3.69 (m, 1H), 3.70-3.77 (m, 1H), 4.33 (dd, J=5.0, 10.0 Hz, 1H),4.40 (dd, J=5.0, 10.0 Hz, 1H), 4.98-5.02 (m, 2H), 5.14 (dd, J=5.0, 15.0Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.12, 16.28, 17.79, 22.61, 22.71,25.95, 27.52, 27.80, 30.74, 30.78, 35.19, 35.25, 40.79, 40.90, 55.57,55.94, 56.87, 57.17, 63.18, 63.37, 121.78, 121.80, 125.24, 125.48,132.09, 136.15, 140.03, 140.11, 171.84, 171.91, 173.35, 173.54, 177.14,177.15; ES-MS: mass calcd for Chemical Formula: C₂₃H₃₈N₂O₅S 454.62.Found (M+) m/z 455.3, (M+Na) m/z 477.3.

Example 26

Synthesis of((R)-2-(3-acetamidopropanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-43)

In a 100 mL round bottom flask, N-acetyl-DL-beta-alanine (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and was stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (250 mg, 57%) to yield Compound N-43. ¹H-NMR (500 MHz, CD₃OD): δ1.50 (s, 6H), 1.57 (s, 3H), 1.60 (s, 3H), 1.83 (s, 3H), 1.86-1.89 (m,2H), 1.95-2.05 (m, 6H), 2.38 (t, J=6.5 Hz, 2H), 2.63 (dd, J=9.0, 14.0Hz, 1H), 2.92 (dd, J=4.0, 14.0 Hz, 1H), 3.06 (dd, J=7.0, 13.0 Hz, 1H),3.21 (m, 1H), 3.36 (m, 2H), 4.50 (dd, J=4.0, 9.0 Hz, 1H), 4.98-5.03 (m,2H), 5.13 (t, J=7.5 Hz, 1H), 5.61 (d, J=10.0 Hz, 1H), 6.17 (d, J=17.0Hz, 1H), 6.28 (dd, J=10.0, 17.0 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ16.16, 16.25, 17.81, 22.66, 25.96, 27.40, 27.79, 30.11, 33.37, 36.41,37.07, 40.80, 40.89, 53.31, 121.56, 125.13, 125.46, 132.12, 136.28,140.58, 173.41, 173.83, 174.01; ES-MS: mass calcd for Chemical Formula:C₂₃H₃₈N₂O₄S 438.62. Found (M+) m/z 439.3, (M+Na) m/z 461.2.

Example 27

Synthesis of a mixture of((R)-2-((R)-2-acetamido-5-amino-5-oxopentanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) and((R)-2-((S)-2-acetamido-5-amino-5-oxopentanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-61)

In a 100 mL round bottom flask, N-acetyl-L-glutamine (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture. The reaction mixture was stirred at roomtemperature overnight. CH₂Cl₂ was removed by rotary evaporation. Theresulting residue was dissolved in ethyl acetate (50 mL). The organicsolution was washed with an NH₄Cl saturated solution (50 mL), dried overNa₂SO₄, and concentrated to afford a crude mixture. The crude mixturewas purified by preparative HPLC (155 mg, 31%) to yield a 1:1 ratiomixture of R-R and S-R isomers of Compound N-61, similar to Compound Cracemate in Examples 5 and 5a. ¹H-NMR (500 MHz, CD₃OD): δ 1.62 (s, 6H),1.69 (s, 3H), 1.70 (s, 3H), 1.98-2.12 (m, 13H), 2.35 (t, J=5.0 Hz, 2H),2.86 (m, 1H), 3.03 (m, 1H), 3.20-3.24 (m, 2H), 4.43-4.46 (m, 2H),5.12-5.13 (m, 2H), 5.26 (m, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.12,16.29, 16.30, 22.54, 22.65, 25.95, 27.54, 27.80, 29.22, 29.25, 30.70,30.78, 32.71, 32.77, 35.35, 35.46, 40.80, 40.90, 54.37, 54.44, 55.38,55.70, 121.77, 121.82, 125.25, 125.48, 132.09, 136.15, 140.01, 140.09,172.78, 172.88, 173.21, 173.34, 177.10, 177.12, 177.99, 178.03; ES-MS:mass calcd for Chemical Formula: C₂₅H₄₁N₃O₅S 495.68. Found (M+) m/z496.4, (M+Na) m/z 518.4.

Example 28

Synthesis of a mixture of((R)-2-((2S,3S)-2-acetamido-3-methylpentanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid and(R)-2-((2R,3S)-2-acetamido-3-methylpentanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-62)

In a 100 mL round bottom flask, N-acetyl-L-isoleucine (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (220 mg, 46%) to yield a mixture of Compound N-62a (the S-S-Renantiomer) and Compound N-62b (the S-R-R enantiomer), wherein the ratioof N-62a to N-62b is 1:1. ¹H-NMR (500 MHz, CD₃OD): δ 0.80-0.86 (m, 6H),1.06-1.33 (m, 3H), 1.50 (s, 6H), 1.57 (s, 3H), 1.60 (s, 3H), 1.79-2.01(m, 11H), 2.71-2.77 (m, 1H), 2.87-2.94 (m, 1H), 3.09-3.11 (m, 2H),4.17-4.43 (m, 2H), 4.99-5.00 (m, 2H), 5.26 (t, J=10.0 Hz, 1H). ¹³C-NMR(125 MHz, CD₃OD): δ 11.68, 12.14, 15.00, 16.13, 16.28, 17.80, 22.52,22.58, 25.86, 25.96, 27.48, 27.52, 27.53, 27.80, 30.80, 30.88, 35.51,35.56, 38.02, 38.31, 40.80, 40.90, 55.37, 55.62, 57.97, 59.80, 121.83,121.88, 125.23, 125.48, 132.08, 136.14, 139.92, 140.02, 172.89, 173.01,173.39, 173.46, 176.84, 177.09; ES-MS: mass calcd for Chemical Formula:C₂₆H₄₄N₂O₄S 480.70. Found (M+) m/z 481.4, (M+Na) m/z 503.4.

Example 29

Synthesis of((R)-2-((S)-5-(benzyloxy)-2-(tert-butoxycarbonylamino)-5-oxopentanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-63)

In a 100 mL round bottom flask, N-Boc-L-glutamic acid-benzyl-ester (1.0mmol), coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (182 mg, 28%) to yield Compound N-63. ¹H-NMR (500 MHz, CDCl₃): δ1.44 (s, 9H), 1.62 (s, 6H), 1.67 (s, 3H), 1.70 (s, 3H), 1.97-2.07 (m,10H), 2.52-2.55 (m, 2H), 2.90 (dd, J=5.0, 15.0 Hz, 1H), 3.01 (dd, J=5.0,15.0 Hz, 1H), 3.18-3.25 (m, 2H), 4.32 (dd, J=10.0, 15.0 Hz, 1H), 4.74(m, 1H), 5.11-5.12 (m, 2H), 5.14 (s, 2H), 5.23 (t, J=10.0 Hz, 1H), 5.50(d, J=10.0 Hz, 1H), 7.22 (d, J=10.0 Hz, 1H), 7.34-7.38 (m, 5H). ¹³C-NMR(125 MHz, CDCl₃): δ 15.99, 16.18, 17.74, 25.72, 26.50, 26.73, 27.85,28.32, 29.90, 32.84, 39.73, 52.02, 53.61, 66.70, 80.44, 119.49, 123.68,124.31, 128.34, 128.61, 131.38, 135.25, 135.62, 140.01, 155.76, 171.70,173.27, 173.51, 207.33; ES-MS: mass calcd for Chemical Formula:C₃₅H₅₂N₂O₇S 644.86. Found (M+) m/z 645.4, (M+Na) m/z 667.5.

Example 30

Synthesis of a mixture of((R)-3-(tert-butoxycarbonylamino)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid and(S)-3-(tert-butoxycarbonylamino)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid) (Compound N-64)

In a 100 mL round bottom flask, N-Boc-L-glutamic acid (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture and stirred at room temperature overnight.CH₂Cl₂ was removed by rotary evaporation. The resulting residue wasdissolved in ethyl acetate (50 mL). The organic solution was washed withan NH₄Cl saturated solution (50 mL), dried over Na₂SO₄, and concentratedto afford a crude mixture. The crude mixture was purified by preparativeHPLC (136 mg, 17%) to yield a 1:1 ratio mixture of R-R and S-R isomersof Compound N-64, similar to Compound C racemate in Examples 5 and 5a.¹H-NMR (500 MHz, CDCl₃): δ 1.36 (s, 9H), 1.53 (s, 6H), 1.59 (s, 3H),1.61 (s, 3H), 1.88-2.03 (m, 10H), 2.77-2.86 (m, 1H), 2.94-2.97 (m, 1H),3.11-3.17 (m, 2H), 4.45 (m, 1H), 4.63 (m, 1H), 5.01-5.03 (m, 2H),5.12-5.15 (m, 1H), 5.89-5.90 (m, 1H), 7.33 (m, 1H), 8.70 (broad, 2H).¹³C-NMR (125 MHz, CDCl₃): δ 15.00, 15.13, 16.68, 24.71, 25.43, 25.67,27.29, 28.70, 28.79, 28.85, 31.43, 36.68, 38.62, 38.67, 49.38, 51.36,79.65, 79.85, 118.23, 118.33, 122.69, 123.27, 130.33, 134.31, 134.34,134.37, 139.32, 154.89, 170.77, 172.68, 172.79; ES-MS: mass calcd forChemical Formula: C₂₇H₄₄N₂O₇S 540.71. Found (M+Na) m/z 563.4.

Example 31

Synthesis of a mixture of((R)-2-((S)-2-acetamido-3-(4-hydroxyphenyl)propanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid and(R)-2-((R)-2-acetamido-3-(4-hydroxyphenyl)propanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-65)

In a 100 mL round bottom flask, N-acetyl-DL-tyrosine (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture. The reaction mixture was stirred at roomtemperature overnight. CH₂Cl₂ was removed by rotary evaporation. Theresulting residue was dissolved in ethyl acetate (50 mL). The organicsolution was washed with NH₄Cl saturated solution (50 mL), dried overNa₂SO₄, and concentrated to afford a crude mixture. The crude mixturewas purified by preparative HPLC (230 mg, 43%) to yield a 1:1 ratiomixture of R-R and S-R isomers of Compound N-65, similar to Compound Cracemate in Examples 5 and 5a. ¹H-NMR (500 MHz, CD₃OD): δ 1.50 (s, 6H),1.56 (s, 3H), 1.59 (s, 3H), 1.79-1.80 (m, 3H), 1.87-88 (m, 2H),1.96-2.03 (m, 8H), 2.57-2.80 (m, 1H), 2.87 (m, 1H), 2.96-3.13 (m, 2H),4.43-4.48 (m, 1H), 4.52-4.55 (m, 1H), 5.00-5.01 (m, 2H), 5.11 (m, 1H),6.59 (d, J=8.0 Hz, 2H), 6.97 (d, J=8.0 Hz, 2H). ¹³C-NMR (125 MHz,CD₃OD): δ 16.26, 16.29, 17.82, 22.43, 25.97, 27.42, 27.79, 30.18, 30.29,33.38, 33.43, 38.21, 38.46, 40.80, 40.89, 53.21, 53.49, 56.12, 56.17,116.13, 116.16, 121.55, 121.59, 125.15, 125.47, 129.06, 129.12, 131.33,131.36, 132.11, 136.26, 140.54, 157.24, 157.30, 173.08, 173.65, 173.70,173.82; ES-MS: mass calcd for Chemical Formula: C₂₉H₄₂N₂O₅S 530.72.Found (M+) m/z 531.3, (M+Na) m/z 553.3.

Example 32

Synthesis of a mixture of((R)-2-((S)-2-acetamidopropanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid and(R)-2-((R)-2-acetamidopropanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-40)

In a 100 mL round bottom flask, N-acetyl-DL-alanine (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-Farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture. The reaction mixture was stirred at roomtemperature overnight. CH₂Cl₂ was removed by rotary evaporation. Theresulting residue was dissolved in ethyl acetate (50 mL). The organicsolution was washed with an NH₄Cl saturated solution (50 mL), dried overNa₂SO₄, and concentrated to afford a crude mixture. The crude mixturewas purified by preparative HPLC (220 mg, 50%) to yield a 1:1 ratiomixture of R-R and S-R isomers of Compound N-40, similar to Compound Cracemate in Examples 5 and 5a. ¹H-NMR (500 MHz, CD₃OD): δ 1.27 (t, J=6.5Hz, 3H), 1.50 (s, 6H), 1.57 (s, 3H), 1.59 (s, 3H), 1.86-2.03 (m, 11H),2.62-2.69 (m, 1H), 2.86-2.91 (m, 1H), 3.04-3.05 (m, 1H), 3.15 (m, 1H),4.32-4.34 (m, 1H), 4.46-4.47 (m, 1H), 4.99-5.01 (m, 2H), 5.11 (m, 1H).¹³C-NMR (125 MHz, CD₃OD): δ 16.14, 16.23, 17.79, 18.10, 18.37, 22.42,25.94, 27.41, 27.78, 30.13, 30.28, 33.35, 33.57, 40.79, 40.88, 50.30,53.08, 53.43, 121.57, 121.60, 125.15, 125.45, 132.11, 140.54, 173.75;ES-MS: mass calcd for Chemical Formula: C₂₃H₃₈N₂O₄S 438.62. Found (M+)m/z 439.2.

Example 33

Synthesis of a mixture of((R)-2-((S)-2-acetamido-3-methylbutanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid and(R)-2-((R)-2-acetamido-3-methylbutanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-41)

In a 100 mL round bottom flask, N-acetyl-DL-valine (1.0 mmol), couplingreagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-Farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture. The reaction mixture was stirred at roomtemperature overnight. CH₂Cl₂ was removed by rotary evaporation. Theresulting residue was dissolved in ethyl acetate (50 mL). The organicsolution was washed with an NH₄Cl saturated solution (50 mL), dried overNa₂SO₄, and concentrated to afford a crude mixture. The crude mixturewas purified by preparative HPLC (250 mg, 54%) to yield a 1:1 ratiomixture of R-R and S-R isomers of Compound N-41, similar to Compound Cracemate in Examples 5 and 5a. ¹H-NMR (500 MHz, CD₃OD): δ 1.27 (m, 6H),1.50 (s, 6H), 1.57 (s, 3H), 1.59 (s, 3H), 1.87-2.03 (m, 12H), 2.61-2.67(m, 1H), 2.86-2.89 (m, 1H), 3.05-3.06 (m, 1H), 3.13-3.15 (m, 1H), 4.21(dd, J=6.5, 16.0 Hz, 1H), 4.47-4.48 (m, 1H), 4.99-5.03 (m, 2H), 5.13 (t,J=8.0 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.16, 17.82, 18.45, 18.62,19.75, 19.93, 22.47, 25.97, 27.42, 27.79, 30.09, 30.24, 32.09, 33.28,33.42, 40.80, 40.89, 53.20, 53.43, 60.00, 60.03, 121.56, 121.60, 125.13,125.15, 125.46, 132.11, 136.26, 140.52, 173.25, 173.32, 173.67, 173.71,173.74; ES-MS: mass calcd for Chemical Formula: C₂₅H₄₂N₂O₄S 466.68.Found (M+) m/z 467.3.

Example 34

Synthesis of((S)-4-(tert-butoxycarbonylamino)-5-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-5-oxopentanoicacid) (Compound N-66)

In a 100 mL round bottom flask, to a solution of(S)-5-(benzyloxy)-2-(tert-butoxycarbonylamino)-5-oxopentanoic acid (674mg, 2 mmol) and benzotriazol-1-yl-oxytripyrrolidinophosphoniumhexafluorophosphate (PyBop, 1040 mg, 2 mmol) and in THF (5 mL) was addedN,N-diisopropyl-ethyl-amine (1.04 mL, 6 mmol) dropwise. After 10 min,S-trans, trans-farnesyl-L-cysteine (650 mg, 2 mmol) was added slowly.The solution was stirred at room temperature for 4 h. The reaction wasquenched by 1 N HCl and pH of the solution was adjusted to 3.0. Themixture was extracted by ethyl acetate (15 mL×3). The organic layer wasdried over Na₂SO₄ and concentrated in vacuo. Half of the residue wasused directly for next step to afford crude Compound N-66. To this crudeCompound N-66 obtained above dissolved in MeOH (1 mL), was added 5 NNaOH (2 mL, 10 mmol). The reaction was left at room temperature for 10min. The reaction was quenched by 1 N HCl and pH of the solution wasadjusted to 2.0. The mixture was extracted by ethyl acetate (15 mL×3).The organic layer was dried over Na₂SO₄ and concentrated in vacuo. Theresidue was then further purified by preparative HPLC (164 mg, 30%) toyield Compound N-66. ¹H-NMR (500 MHz, CDCl₃): δ 1.45 (s, 9H), 1.62 (s,6H), 1.68 (s, 3H), 1.70 (s, 3H), 1.95-2.09 (m, 10H), 2.46-2.54 (m, 2H),2.91 (dd, J=5.0, 15.0 Hz, 1H), 3.04 (dd, J=5.0, 15.0 Hz, 1H), 3.17-3.25(m, 2H), 4.56 (dd, J=10.0, 15.0 Hz, 1H), 4.77-4.81 (m, 1H), 5.12 (m,2H), 5.23 (t, J=10.0 Hz, 1H), 5.64 (d, J=10.0 Hz, 1H), 7.66 (d, J=10.0Hz, 1H), 8.40 (broad, 2H). ¹³C-NMR (125 MHz, CDCl₃): δ 16.06, 16.15,17.75, 25.77, 26.52, 26.74, 28.01, 28.33, 29.73, 29.89, 32.60, 39.70,39.74, 52.22, 52.95, 80.91, 119.57, 123.83, 124.35, 131.38, 135.34,140.04, 156.06, 172.18, 174.00, 177.16; ES-MS: mass calcd for ChemicalFormula: C₂₈H₄₆N₂O₇S 554.74. Found (M+Na) m/z 577.4.

Example 35

Synthesis of a mixture of((R)-2-((S)-2-acetamidobutanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid and(R)-2-((R)-2-acetamidobutanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-46)

In a 100 mL round bottom flask, to a solution ofN-acetyl-DL-2-amino-n-butyric acid (174 mg, 1.2 mmol) and4-(4,6-dimethoxy-1,3,5-triazin-2yl)-4-methylmorpholinium chloride(DMTMM, 332 mg, 1.2 mmol) in CH₂Cl₂ (5 mL) was addedN,N-diisopropyl-ethyl-amine (0.52 mL, 3 mmol) dropwise. After 10 min,S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was added slowly.The solution was stirred at room temperature overnight. The mixture wasdiluted with ethyl acetate (60 mL) and washed by 0.5 N HCl (15 mL×1),H₂O (15 mL×2) and brine (15 mL×2). The organic layer was dried overNa₂SO₄ and concentrated in vacuo. The residue was purified bypreparative HPLC (290 mg, 64%) to yield a 1:1 ratio mixture of R-R andS-R isomers of Compound N-46, similar to Compound C racemate in Examples5 and 5a. ¹H-NMR (500 MHz, CD₃OD): δ 0.86 (m, 3H), 1.50 (s, 6H), 1.57(s, 3H), 1.60 (s, 3H), 1.72-2.03 (m, 13H), 2.61-2.68 (m, 1H), 2.87-2.89(m, 1H), 3.04 (m, 1H), 3.12-3.21 (m, 1H), 4.21-4.25 (m, 1H), 4.45-4.48(m, 1H), 4.99-5.01 (m, 2H), 5.13 (t, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz,CD₃OD): δ 10.62, 10.70, 16.16, 16.25, 17.81, 22.46, 25.96, 26.56, 26.68,27.42, 27.79, 30.11, 30.27, 33.31, 33.52, 40.80, 40.89, 53.11, 53.42,56.02, 56.07, 121.56, 121.60, 125.13, 125.46, 132.11, 136.26, 136.27,140.54, 173.28, 173.32, 173.69, 174.26, 174.31; ES-MS: mass calcd forChemical Formula: C₂₄H₄₀N₂O₄S 452.65. Found (M+) m/z 453.3, (M+Na) m/z475.2.

The following general experimental procedures for loadingFmoc-cysteine-(S-farnesyl) on resin were used for Examples 36-39 asdescribed below. 2-Chlorotrityl chloride resin (loading efficiency=1.01mmol/g, 1.0 g, 1.0 mmol) was placed in a 50 mL peptide synthesis vessel(Polypropylene syringe from Torviq, Niles, Mich.) under Nitrogen. Tothis was added anhydrous CH₂Cl₂ (20 mL). The resin was shacked in 5 minand solvent was removed. In a separate vial, Fmoc-Cys(StBu)-OH (1.1 g,2.6 mmol) and 2,4,6-collidine (290 mg, 2.8 mmol) were dissolved inanhydrous CH₂Cl₂ (20 mL). This solution was transferred to the resin.The mixture was gently agitated for 3 h. Then, 1% solution of2,4,6-collidine in MeOH (20 mL) was then added, and the mixture wasagitated for an additional 10 min. The mixture was drained, and theresin was washed MeOH, CH₂Cl₂, and DMF. Dithiothreitol (1.1 g, 6.7 mmol)was dissolved in a diisopropylethylamine/DMF solution (4 mL/20 mL) andadded to the resin, and the reaction vessel was gently agitatedovernight. The solvent was drained, and the resin washed with CH₂Cl₂,and DMF. 2,4,6-Collidine (300 mg, 2.5 mmol) was added to a solution offarnesyl bromide (900 mg, 3.2 mmol) in CH₂Cl₂ (20 mL). This reagentsolution was added to the resin, and the reaction vessel was gentlyagitated for 10 h at room temperature. The solvent was then drained, andthe resin was washed with DMF and CH₂Cl₂.

Example 36

Synthesis of((R)-2-((2S,3S)-3-methyl-2-(methylsulfonamido)pentanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-67)

In a 100 mL round bottom flask, 20% solution of piperidine in DMF (10mL) was added to the farnesylcysteine on the resin (0.5 mmol) and thevessel was agitated for 15 min. The resin was washed with DMF andCH₂Cl₂. Fmoc-L-isoleucine (354 mg, 1 mmol), PBOP (502 mg, 1 mmol) and2,4,6-collidine (242 mg, 2 mmol) were dissolved in DMF (5 mL). Thereaction mixture was stirred in 5 min. This solution was added to theresin and agitated for 3 h. The solvent was then drained, and the resinwas washed with DMF and CH₂Cl₂. 20% solution of piperidine in DMF (10mL) was added to the farnesylcysteine on the resin (0.5 mmol) and thevessel was agitated for 15 min. The resin was washed with DMF andCH₂Cl₂. Methyl sulfonyl chloride (1 mmol) and 2,4,6-collidine (242 mg, 2mmol) were dissolved in DMF (5 mL). This solution was added to the resinand agitated for 3 h. The solvent was then drained, and the resin waswashed with DMF and CH₂Cl₂. The resin was treated twice with a 0.5%solution of trifluoroacetic acid in CH₂Cl₂ for 5 min. The solution wascollected into a round-bottom flask, and the resin was washed twice withanhydrous CH₂Cl₂. The solvent was removed by rotary evaporation. Theproduct was purified by preparative HPLC (45 mg, 25%) to yield CompoundN-67. ¹H-NMR (500 MHz, CD₃OD): δ 0.94 (t, J=5.0 Hz, 3H), 1.02 (d, J=5.0Hz, 3H), 1.22 (m, 2H), 1.62 (s, 3H), 1.66 (s, 3H), 1.67 (s, 3H), 1.70(s, 3H), 1.77-1.82 (m, 1H), 1.99 (t, J=7 Hz, 2H), 2.06-2.17 (m, 6H),2.74 (dd, J=8.0, 14.0 Hz, 1H), 2.97 (S, 3H), 3.08 (dd, J=5.0, 12.5 Hz,1H), 3.15 (dd, J=7.0, 13.0 Hz, 1H), 3.30 (dd, J=5.0, 12.0 Hz, 1H), 3.81(d, J=5.0 Hz, 1H), 4.62 (dd, J=5.0, 10.0 Hz, 1H), 5.10-5.16 (m, 2H),5.24 (t, J=7.0 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 11.3, 16.0, 16.3,25.7, 26.0, 27.4, 27.8, 39.2, 40.8, 40.9, 41.3, 52.2, 62.8, 121.5,125.1, 125.5, 132.1, 136.3, 140.6, 173.6, 174.0; ES-MS: mass calcd forChemical Formula: C₂₅H₄₄N₂O₅S₂ 516.8. Found (M+) m/z 517.

Example 37

Synthesis of((R)-2-((2S,3S)-3-methyl-2-(phenylsulfonamido)pentanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-68)

In a 100 mL round bottom flask, 20% solution of piperidine in DMF (10mL) was added to the farnesylcysteine on the resin (0.5 mmol) and thevessel was agitated for 15 min. The resin was washed with DMF andCH₂Cl₂. Fmoc-L-isoleucine (354 mg, 1 mmol), PBOP (502 mg, 1 mmol) and2,4,6-collidine (242 mg, 2 mmol) were dissolved in DMF (5 mL). Thereaction mixture was stirred in 5 min. This solution was added to theresin and agitated for 3 h. The solvent was then drained, and the resinwas washed with DMF and CH₂Cl₂. 20% solution of piperidine in DMF (10mL) was added to the farnesylcysteine on the resin (0.5 mmol) and thevessel was agitated for 15 min. The resin was washed with DMF andCH₂Cl₂. Phenyl sulfonyl chloride (1 mmol) and 2,4,6-collidine (242 mg, 2mmol) were dissolved in DMF (5 mL). This solution was added to the resinand agitated for 3 h. The solvent was then drained, and the resin waswashed with DMF and CH₂Cl₂. The resin was treated twice with a 0.5%solution of trifluoroacetic acid in CH₂Cl₂ for 5 min. The solution wascollected into a round-bottom flask, and the resin was washed twice withanhydrous CH₂Cl₂. The solvent was removed by rotary evaporation. Theproduct was purified by preparative HPLC (40 mg, 30%) to yield CompoundN-68. ¹H-NMR (500 MHz, CD₃OD): δ 0.74 (t, J=7.5 Hz, 3H), 0.80 (d, J=7.0Hz, 3H), 1.00-1.06 (m, 2H), 1.43-1.48 (m, 1H), 1.50 (s, 3H), 1.51 (s,3H), 1.57 (s, 3H), 1.58 (s, 3H), 1.87 (t, J=7 Hz, 2H), 1.97-2.07 (m,6H), 2.45 (dd, J=7.5, 13.5 Hz, 1H), 2.62 (dd, J=7.5, 14.0 Hz, 1H), 2.99(dd, J=8.0, 13.0 Hz, 1H), 3.06 (dd, J=8.0, 13.0 Hz, 1H), 3.60 (d, J=7.0Hz, 1H), 4.12 (dd, J=5.5, 7.5 Hz, 1H), 4.99-5.02 (m, 2H), 5.10 (t, J=7.0Hz, 1H), 7.40-7.50 (m, 3H), 7.73 (d, J=8.0 Hz, 2H). ¹³C-NMR (125 MHz,CD₃OD): δ 11.4, 15.8, 16.2, 16.3, 17.8, 25.5, 26.0, 27.4, 27.8, 30.4,33.3, 39.4, 40.8, 40.9, 53.4, 62.4, 121.6, 125.2, 125.5, 128.3, 130.1,132.1, 133.6, 136.3, 140.5, 142.2, 173.0, 173.5; ES-MS: mass calcd forChemical Formula: C₃₀H₄₆N₂O₅S₂ 578.8. Found (M+) m/z 579.3.

Example 38

Synthesis of((R)-2-((2S,3S)-2-(cyclopropanesulfonamido)-3-methylpentanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-69)

In a 100 mL round bottom flask, 20% solution of piperidine in DMF (10mL) was added to the farnesylcysteine on the resin (0.5 mmol) and thevessel was agitated for 15 min. The resin was washed with DMF andCH₂Cl₂. Fmoc-L-isoleucine (354 mg, 1 mmol), PBOP (502 mg, 1 mmol) and2,4,6-collidine (242 mg, 2 mmol) were dissolved in DMF (5 mL). Thereaction mixture was stirred in 5 min. This solution was added to theresin and agitated for 3 h. The solvent was then drained, and the resinwas washed with DMF and CH₂Cl₂. 20% solution of piperidine in DMF (10mL) was added to the farnesylcysteine on the resin (0.5 mmol) and thevessel was agitated for 15 min. The resin was washed with DMF andCH₂Cl₂. Cyclopropyl sulfonyl chloride (1 mmol) and 2,4,6-collidine (242mg, 2 mmol) were dissolved in DMF (5 mL). This solution was added to theresin and agitated for 3 h. The solvent was then drained, and the resinwas washed with DMF and CH₂Cl₂. The resin was treated twice with a 0.5%solution of trifluoroacetic acid in CH₂Cl₂ for 5 min. The solution wascollected into a round-bottom flask, and the resin was washed twice withanhydrous CH₂Cl₂. The solvent was removed by rotary evaporation. Theproduct was purified by preparative HPLC (40 mg, 26%) to yield CompoundN-69. ¹H-NMR (500 MHz, CD₃OD): δ 0.81 (t, J=7.5 Hz, 3H), 0.86 (d, J=6.5Hz, 3H), 1.05-1.11 (m, 2H), 1.50 (bs, 6H), 1.42-1.52 (m, 4H), 1.57 (s,3H), 1.59 (s, 3H), 1.72-1.78 (m, 1H), 1.89 (t, J=7 Hz, 2H), 1.94-2.04(m, 6H), 2.64 (dd, J=8.0, 13.0 Hz, 1H), 2.87 (dd, J=5.0, 13.0 Hz, 1H),3.04 (dd, J=7.5, 13.0 Hz, 1H), 3.14 (dd, J=8.5, 13.5 Hz, 1H), 4.20 (d,J=7.5 Hz, 1H), 4.47 (dd, J=5.0, 8.5 Hz, 1H), 4.98-5.03 (m, 2H), 5.11 (t,J=7.5 Hz, 1H), 7.40-7.50 (m, 3H), 7.73 (d, J=8.0 Hz, 2H). ¹³C-NMR (125MHz, CD₃OD): δ 11.5, 15.9, 16.2, 16.3, 17.8, 22.5, 25.8, 26.0, 27.4,27.8, 30.3, 33.3, 38.3, 40.8, 40.9, 53.5, 59.2, 121.6, 125.2, 125.5,132.1, 136.3, 140.5, 173.2, 173.7, 173.8; ES-MS: mass calcd for ChemicalFormula: C₂₇H₄₆N₂O₅S₂ 542.8. Found (M+) m/z 543.3.

Example 39

Synthesis of((R)-2-((2S,3S)-3-methyl-2-(morpholine-4-carboxamido)pentanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-70)

In a 100 mL round bottom flask, 20% solution of piperidine in DMF (10mL) was added to the farnesylcysteine on the resin (0.5 mmol) and thevessel was agitated for 15 min. The resin was washed with DMF andCH₂Cl₂. Fmoc-L-isoleucine (354 mg, 1 mmol), PBOP (502 mg, 1 mmol) and2,4,6-collidine (242 mg, 2 mmol) were dissolved in DMF (5 mL). Thereaction mixture was stirred in 5 min. This solution was added to theresin and agitated for 3 h. The solvent was then drained, and the resinwas washed with DMF and CH₂Cl₂. 20% solution of piperidine in DMF (10mL) was added to the farnesylcysteine on the resin (0.5 mmol) and thevessel was agitated for 15 min. The resin was washed with DMF andCH₂Cl₂. 4-Morpholine carbonyl chloride (1 mmol) and 2,4,6-collidine (242mg, 2 mmol) were dissolved in DMF (5 mL). This solution was added to theresin and agitated for 3 h. The solvent was then drained, and the resinwas washed with DMF and CH₂Cl₂. The resin was treated twice with a 0.5%solution of trifluoroacetic acid in CH₂Cl₂ for 5 min. The solution wascollected into a round-bottom flask, and the resin was washed twice withanhydrous CH₂Cl₂. The solvent was removed by rotary evaporation. Theproduct was purified by preparative HPLC (40 mg, 30%) to yield CompoundN-70. ¹H-NMR (500 MHz, CD₃OD): δ 0.81 (t, J=7.5 Hz, 3H), 0.86 (d, J=6.5Hz, 3H), 1.05-1.13 (m, 2H), 1.50 (bs, 6H), 1.57 (s, 3H), 1.59 (s, 3H),1.72-1.78 (m, 1H), 1.87 (t, J=7 Hz, 2H), 1.94-2.04 (m, 6H), 2.66 (dd,J=7.5, 13.5 Hz, 1H), 2.88 (dd, J=5.0, 14.0 Hz, 1H), 3.05 (dd, J=7.0,13.5 Hz, 1H), 3.14 (dd, J=8.0, 13.0 Hz, 1H), 3.30 (t, J=5.0 Hz, 2H),3.55 (t, J=5.0 Hz, 2H), 4.07 (d, J=8.0 Hz, 1H), 4.47 (dd, J=5.0, 8.0 Hz,1H), 4.98-5.03 (m, 2H), 5.12 (t, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz,CD₃OD): δ 11.4, 16.0, 16.2, 16.3, 17.8, 26.0, 26.1, 27.4, 27.8, 30.4,33.6, 38.2, 40.8, 40.9, 45.5, 53.5, 67.6, 121.6, 125.2, 125.5, 132.1,136.3, 140.5, 159.6, 173.8, 174.9; ES-MS: mass calcd for ChemicalFormula: C29H49N3O5S 551.8. Found (M+) m/z 552.4.

Example 40

Synthesis of(4-((R)-1-hydrazinyl-1-oxo-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propan-2-ylamino)-4-oxobutanoicacid) (Compound N-24)

In a 100 mL round bottom flask, to a solution of S-trans,trans-farnesyl-L-cysteine methyl ester (430 mg, 1.27 mmol) in THF (10mL) was added succinic anhydride (635 mg, 6.34 mmol). The solution wasstirred at room temperature overnight. The solvent was removed in vacuoand the residue was dissolved with ethyl acetate (60 mL). The solutionwas washed by H₂O (15 mL×2) and brine (15 mL×1). The organic layer wasdried over Na₂SO₄ and concentrated in vacuo. To the residue obtainedabove was added hydrazine in THF (1 M hydrazine in THF, 25 mL). Thereaction was left at room temperature for 24 h and the solvent wasremoved in vacuo. The residue was then further purified by preparativeHPLC (122 mg, 22%) to yield Compound N-24. ¹H-NMR (500 MHz, DMSO-d₆): δ1.55 (s, 6H), 1.62 (s, 3H), 1.64 (s, 3H), 1.91-2.06 (m, 8H), 2.36-2.41(m, 4H), 2.71 (dd, J=5.0, 10.0 Hz, 1H), 3.13-3.17 (m, 3H), 4.39 (dd,J=5.0, 10.0 Hz, 1H), 5.07 (m, 2H), 5.16 (t, J=10.0 Hz, 1H), 8.18 (d,J=10.0 Hz, 1H), 9.27 (s, 1H). ¹³C-NMR (125 MHz, DMSO-d₆): δ 15.75,15.79, 17.55, 25.50, 25.85, 26.14, 28.63, 29.17, 29.89, 32.68, 39.90,39.99, 50.93, 120.15, 123.64, 124.10, 130.65, 134.54, 138.40, 169.30,170.86, 173.96; ES-MS: mass calcd for Chemical Formula: C₂₂H₃₇N₃O₄S439.61. Found (M+) m/z 440.3, (M+Na) m/z 462.2.

Example 41

Synthesis of a mixture of(N-[1-Hydrazinocarbonyl-2-(3,7,11-trimethyl-dodeca-2,6,10-trienylsulfanyl)-ethyl]-3-methyl-succinamicacid) (Compound N-34) and(N-[1-Hydrazinocarbonyl-2-(3,7,11-trimethyl-dodeca-2,6,10-trienylsulfanyl)-ethyl]-2-methyl-succinamicacid) (Compound N-33)

In a 100 mL round bottom flask, S-trans, trans-farnesyl-L-cysteinemethyl ester (339 mg, 1 mmol) and methylsuccinic anhydride (171 mg, 1.5mmol) were mixed in CH₂Cl₂ (5 mL). N,N-diisopropyl-ethyl-amine (0.52 mL,3 mmol) was added to reaction mixture. The reaction solution was stirredat room temperature overnight. Ethyl acetate (50 mL) was added and thenwashed with saturated ammonium chloride aqueous solution (20 mL×2), DIwater (20 mL×2) and brine (20 mL×2) sequentially. The ethyl acetatesolution was dried by Na₂SO₄ and concentrated in vacuo to yield a crudemixture. The crude mixture was purified by HPLC to yield two fractions.The first fraction was purified as explained in Example 41a. The secondfraction collected afforded a 6:4 mixture of the 3-methyl (CompoundN-34) and 2-methyl (Compound N-33) regioisomers, wherein eachregioisomer is a 1:1 ratio mixture of R-R and S-R isomers (150 mg, 33%).¹H-NMR (500 MHz, MeOH-d₄): δ 1.04-1.13 (m, 3H), 1.50 (s, 6H), 1.57 (s,3H), 1.59 (s, 3H), 1.85-1.88 (m, 2H), 1.93-2.03 (m, 6H), 2.21-2.33 (m,1H), 2.48-2.63 (m, 2H), 2.68-2.82 (m, 2H), 3.05-3.14 (m, 2H), 4.32-4.42(m, 1H), 4.98-5.02 (m, 1H), 5.12 (t, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz,MeOH-d₄): δ 16.14, 16.25, 16.27, 17.41, 17.47, 17.80, 17.99, 18.33,20.05, 23.10, 23.66, 26.30, 26.56, 26.85, 27.43, 27.79, 29.09, 30.12,30.17, 30.20, 30.82, 33.37, 33.39, 33.50, 33.53, 33.61, 37.40, 37.59,37.96, 38.03, 38.29, 38.58, 38.84, 39.76, 39.95, 40.08, 40.76, 40.89,121.23, 121.41, 121.44, 125.14, 132.13, 136.25, 140.54, 140.62, 172.06,173.93, 174.29, 176.47, 178.93, 179.30; ES-MS: mass calcd for ChemicalFormula: C₂₃H₃₉N₃O₄S 453.3. Found (M+) m/z 454.3.

Example 41a

Synthesis of a mixture of(N-[1-Hydrazinocarbonyl-2-(3,7,11-trimethyl-dodeca-2,6,10-trienylsulfanyl)-ethyl]-3-(S)-methyl-succinamicacid) and(N-[1-Hydrazinocarbonyl-2-(3,7,11-trimethyl-dodeca-2,6,10-trienylsulfanyl)-ethyl]-3-(R)-methyl-succinamicacid) (Compound N-34)

In a 100 mL round bottom flask, S-trans, trans-farnesyl-L-cysteinemethyl ester (339 mg, 1 mmol) and methylsuccinic anhydride (171 mg, 1.5mmol) were mixed in CH₂Cl₂ (5 mL). N,N-diisopropyl-ethyl-amine (0.52 mL,3 mmol) was added to reaction mixture. The reaction solution was stirredat room temperature overnight. Ethyl acetate (50 mL) was added and thenwashed with saturated ammonium chloride aqueous solution (20 mL×2), DIwater (20 mL×2) and brine (20 mL×2) sequentially. The ethyl acetatesolution was dried by Na₂SO₄ and concentrated in vacuo to yield a crudemixture. The crude mixture was purified by HPLC to yield two fractions.The second fraction was purified to afford the mixture of regioisomersas explained in Example 41. The first fraction was isolated to yieldCompound N-34 (50 mg, 11%): ¹H-NMR (500 MHz, MeOH-d₄): δ 1.05 (d, J=7.0Hz, 3H), 1.09 (d, J=7.0 Hz, 3H), 1.50 (s, 6H), 1.57 (s, 3H), 1.59 (s,3H), 1.85-1.88 (m, 2H), 1.94-2.03 (m, 6H), 2.23-2.29 (m, 1H), 2.50-2.64(m, 2H), 2.70-2.80 (m, 2H), 3.07-3.11 (m, 2H), 4.30-4.34 (m, 1H),4.98-5.02 (m, 1H), 5.12 (t, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz, MeOH-d₄): δ16.13, 16.24, 17.42, 17.79, 17.98, 25.94, 27.43, 27.45, 27.96, 27.79,30.19, 30.39, 33.36, 33.49, 37.45, 37.99, 38.91, 39.97, 40.76, 40.89,53.04, 53.34, 121.41, 121.93, 125.14, 125.15, 125.45, 132.12, 136.24,140.62, 172.06, 173.95, 178.17; ES-MS: mass calcd for Chemical Formula:C₂₃H₃₉N₃O₄S 453.3. Found (M+) m/z 454.3.

Example 42

Synthesis of(N1-((R)-1-hydrazinyl-1-oxo-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propan-2-yl)succinamide)(Compound N-38)

In a 100 mL round bottom flask, Succinamic acid (140 mg, 1.2 mmol),2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (1.1 mg, 1.1 mmol) and N,N-diisopropyl-ethyl-amine(0.52 mL, 3 mmol) were mixed in THF (5 mL). The reaction solution wasstirred at room temperature for ten minutes. S-trans,trans-Farnesyl-L-cysteine methyl ester (339 mg, 1 mmol) was added toreaction mixture. The reaction solution was stirred at room temperatureovernight. Ethyl acetate (50 mL) was added and then washed withsaturated ammonium chloride aqueous solution (20 mL×2), DI water (20mL×2) and brine (20 mL×2) sequentially. The ethyl acetate solution wasdried by Na₂SO₄ and concentrated in vacuo to afford a crude mixture, Thecrude mixture obtained was added to 1M NH₂NH₂ in THF (10 mL, 10 mmol).The reaction solution was stirred at room temperature overnight. The THFsolution was concentrated in vacuo to afford a crude mixture. The crudemixture was purified by HPLC (153 mg, 35%) to yield Compound N-38.¹H-NMR (500 MHz, MeOH-d₄): δ 1.50 (s, 6H), 1.57 (s, 3H), 1.59 (s, 3H),1.85-1.88 (m, 2H), 1.93-1.98 (m, 4H), 1.99-2.03 (m, 2H), 2.36-2.49 (m,4H), 2.71 (dd, J=7.5, 13.5 Hz, 1H), 2.83 (dd, J=5.5, 13.5 Hz, 1H), 3.09(d, J=8.0 Hz, 2H), 4.37 (t, J=7.5 Hz, 1H), 5.00 (m, 2H), 5.13 (t, J=8.0Hz, 1H). ¹³C-NMR (125 MHz, MeOH-d₄): δ 16.13, 16.24, 17.79, 25.94,27.42, 27.79, 30.22, 31.34, 31.87, 33.59, 40.76, 40.88, 53.19, 121.44,125.14, 125.46, 132.12, 136.25, 140.58, 171.96, 174.95, 177.38; ES-MS:mass calcd for Chemical Formula: C₂₂H₃₈N₄O₃S 438.3. Found (M+) m/z439.3.

Example 43

Synthesis of((R)-2-(4-methoxy-4-oxobutanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-17)

In a 100 mL round bottom flask, mono-methyl succinate (132 mg, 1 mmol),2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (1.1 mg, 1.1 mmol) and N,N-diisopropyl-ethyl-amine(0.52 mL, 3 mmol) were mixed in THF (5 mL). The reaction solution wasstirred at room temperature for ten minutes. S-trans,trans-Farnesyl-L-cysteine (325 mg, 1 mmol) was added to reactionmixture. The reaction solution was stirred at room temperatureovernight. Ethyl acetate (50 mL) was added and then washed withsaturated ammonium chloride aqueous solution (20 mL×2), DI water (20mL×2) and brine (20 mL×2) sequentially. The ethyl acetate solution wasdried by Na₂SO₄ and concentrated in vacuo to afford a crude CompoundN-17. The crude Compound N-17 was purified by HPLC (110 mg, 25%) toyield Compound N-17. ¹H-NMR (500 MHz, MeOH-d₄): δ 1.50 (s, 6H), 1.57 (s,3H), 1.58 (s, 3H), 1.85-1.88 (m, 2H), 1.91-1.96 (m, 4H), 1.99-2.03 (m,2H), 2.46-2.54 (m, 4H), 2.68 (dd, J=7.5, 13.5 Hz, 1H), 2.90 (dd, J=4.5,13.5 Hz, 1H), 3.09-3.12 (m, 2H), 3.21 (s, 3H), 4.35 (dd, J=4.5, 7.0 Hz,1H), 4.98-5.02 (m, 2H), 5.14 (t, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz,MeOH-d₄): δ 16.12, 16.25, 17.79, 25.94, 27.48, 27.79, 30.40, 30.69,31.73, 35.49, 40.77, 40.89, 52.24, 55.45, 121.82, 125.22, 125.47,132.08, 136.15, 139.97, 173.57, 174.80, 177.17; ES-MS: mass calcd forChemical Formula: C₂₃H₃₇NO₅S 439.2. Found (M+Na) m/z 462.2.

Example 44

Synthesis of ((methyl4-((R)-1-hydrazinyl-1-oxo-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propan-2-ylamino)-4-oxobutanoate))(Compound N-44)

In a 100 mL round bottom flask, the crude Compound N-39 (1 mmol) ofExample 43, -(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexaflurophosphate (1.1 mg, 1.1 mmol) and N,N-diisopropyl-ethyl-amine(0.52 mL, 3 mmol) were mixed in THF (5 mL). The reaction solution wasstirred at room temperature for ten minutes. 1 N Hydrazine in THF (2 mL,2 mmol) was added to reaction mixture. The reaction solution was stirredat room temperature overnight. Ethyl acetate (50 mL) was added and thenwashed with saturated ammonium chloride aqueous solution (20 mL×2), DIwater (20 mL×2) and brine (20 mL×2) sequentially. The ethyl acetatesolution was dried by Na₂SO₄ and concentrated in vacuo to afford a crudemixture, The crude mixture was purified by HPLC (30 mg, 26%) to yieldCompound N-44. ¹H-NMR (500 MHz, MeOH-d₄): δ 1.50 (s, 6H), 1.57 (s, 3H),1.59 (s, 3H), 1.85-1.88 (m, 2H), 1.93-1.98 (m, 4H), 1.99-2.03 (m, 2H),2.42-2.45 (m, 2H), 2.52-2.54 (m, 2H), 2.54-2.59 (m, 1H), 2.80 (dd,J=6.5, 13.5 Hz, 1H), 3.09 (d, J=8 Hz, 2H), 3.57 (s, 3H), 4.37 (t, J=6.0Hz, 1H), 4.98-5.01 (m, 2H), 5.12 (t, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz,MeOH-d₄): δ 16.14, 16.25, 17.80, 25.95, 27.43, 27.79, 30.07, 30.20,31.24, 33.60, 40.76, 40.89, 52.34, 53.08, 121.43, 125.14, 125.46,132.13, 136.25, 140.60, 172.00, 174.35, 174.98; ES-MS: mass calcd forChemical Formula: C₂₃H₃₉N₃O₄S 453.3. Found (M+Na) m/z 476.2.

Example 45

Synthesis of(4-hydrazinyl-N—((R)-1-hydrazinyl-1-oxo-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propan-2-yl)-4-oxobutanamide)(Compound N-25)

In a 100 mL round bottom flask, to a solution of S-trans,trans-farnesyl-L-cysteine methyl ester (339 mg, 1 mmol),benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate(PyBop, 624 mg, 1.2 mmol) and 4-methoxy-4-oxobutanoic acid (1.2 mmol) inTHF (5 mL) was added N,N-diisopropyl-ethyl-amine (0.52 mL, 3 mmol). Thesolution was stirred at room temperature overnight. The mixture wasdiluted with ethyl acetate (60 mL) and washed by 0.5 N HCl (10 mL×1),H₂O (15 mL×2) and brine (10 mL×1). The organic layer was dried overNa₂SO₄ and concentrated in vacuo. The residue was purified by flashchromatography on silica gel with hexanes/ethyl acetate (3/1) as eluent.To the product obtained above was added hydrazine in THF (1 M hydrazinein THF, 48 mL). The reaction was left at room temperature for 64 h andthe solvent was removed in vacuo. The residue was then further purifiedby preparative HPLC (300 mg, 68%) to yield Compound N-25. ¹H-NMR (500MHz, CD₃OD): δ 1.50 (s, 6H), 1.57 (s, 3H), 1.59 (s, 3H), 1.85-2.03 (m,8H), 2.35-2.46 (m, 4H), 2.60 (dd, J=10.0, 15.0 Hz, 1H), 2.87 (dd, J=5.0,15.0 Hz, 1H), 3.08-3.10 (m, 2H), 4.38 (dd, J=5.0, 10.0 Hz, 1H),4.99-5.01 (m, 2H), 5.16 (t, J=10.0 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ16.14, 16.26, 17.80, 25.95, 27.44, 27.80, 30.06, 30.24, 31.96, 33.58,40.77, 40.89, 53.28, 121.44, 125.15, 125.47, 132.13, 136.27, 140.59,171.98, 173.96, 174.88; ES-MS: mass calcd for Chemical Formula:C₂₂H₃₉N₅O₃S 453.64. Found (M+) m/z 454.3.

Example 46

Synthesis of((S)-3-acetamido-4-hydrazinyl-N—((R)-1-hydrazinyl-1-oxo-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propan-2-yl)-4-oxobutanamide)(Compound N-71)

In a 100 mL round bottom flask, to a solution of S-trans,trans-farnesyl-L-cysteine methyl ester (339 mg, 1 mmol),benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate(PyBop, 624 mg, 1.2 mmol) and N-acetyl-aspartic acid methyl ester (1.2mmol) in THF (5 mL) was added N,N-diisopropyl-ethyl-amine (0.52 mL, 3mmol). The solution was stirred at room temperature overnight. Themixture was diluted with ethyl acetate (60 mL) and washed by 0.5 N HCl(10 mL×1), H₂O (15 mL×2) and brine (10 mL×1). The organic layer wasdried over Na₂SO₄ and concentrated in vacuo. The residue was purified byflash chromatography on silica gel with hexanes/ethyl acetate (3/1) aseluent. To the product obtained above was added hydrazine in THF (1 Mhydrazine in THF, 48 mL). The reaction was left at room temperature for64 h and the solvent was removed in vacuo. The residue was then furtherpurified by preparative HPLC (330 mg, 65%) to yield Compound N-71.¹H-NMR (500 MHz, DMSO-d₆): δ 1.56 (s, 6H), 1.62 (s, 3H), 1.63 (s, 3H),1.81 (s, 3H), 1.91-2.04 (m, 10H), 2.38 (dd, J=10.0, 15.0 Hz, 1H),2.68-2.72 (m, 1H), 3.12-3.14 (m, 2H), 4.20 (s, 2H), 4.28 (s, 2H),4.35-4.37 (m, 1H), 4.51-4.52 (m, 1H), 5.06-5.07 (m, 2H), 5.15 (t, J=10.0Hz, 1H), 7.95 (d, J=10.0 Hz, 1H), 8.13 (d, J=10.0 Hz, 1H), 9.10 (s, 1H),9.28 (s, 1H). ¹³C-NMR (125 MHz, DMSO-d₆): δ 15.78, 17.56, 22.67, 25.50,25.88, 26.14, 28.63, 32.53, 37.80, 48.72, 50.99, 120.07, 123.66, 124.09,130.65, 134.53, 138.45, 168.94, 168.98, 169.08, 170.10; ES-MS: masscalcd for Chemical Formula: C₂₄H₄₂N₆O₄S 510.69. Found (M+) m/z 511.3.

Example 47

Synthesis of a mixture of ((2R,3S)-2-acetamido-N—((R)-1-hydrazinyl-1-oxo-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propan-2-yl)-3-methylpentanamide)and ((2R,3R)-2-acetamido-N—((R)-1-hydrazinyl-1-oxo-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propan-2-yl)-3-methylpentanamide)(Compound N-72)

In a 100 mL round bottom flask, to a solution of S-trans,trans-farnesyl-L-cysteine methyl ester (339 mg, 1 mmol),benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate(PyBop, 624 mg, 1.2 mmol) and N-acetyl-L-isoleucine (1.2 mmol) in THF (5mL) was added N,N-diisopropyl-ethyl-amine (0.52 mL, 3 mmol). Thesolution was stirred at room temperature overnight. The mixture wasdiluted with ethyl acetate (60 mL) and washed by 0.5 N HCl (10 mL×1),H₂O (15 mL×2) and brine (10 mL×1). The organic layer was dried overNa₂SO₄ and concentrated in vacuo. The residue was purified by flashchromatography on silica gel with hexanes/ethyl acetate (3/1) as eluent.To the product obtained above was added hydrazine in THF (1 M hydrazinein THF, 48 mL). The reaction was left at room temperature for 64 h andthe solvent was removed in vacuo. The residue was then further purifiedby preparative HPLC (245 mg, 50%) to yield a 1:1 ratio mixture of RSRand RRR isomers of Compound N-72, similar to the Compound N-62 racematein Example 28, wherein one of the three chiral centers is racemic andthe other two are enantiopure. ¹H-NMR (500 MHz, CD₃OD): δ 0.79-0.87 (m,6H), 1.05-1.15 (m, 2H), 1.50-1.59 (m, 13H), 1.85-2.01 (m, 11H), 2.57 (m,1H), 2.75-2.94 (m, 1H), 3.07-3.10 (m, 2H), 4.07-4.16 (m, 1H), 4.36-4.37(m, 1H), 4.99-5.01 (m, 2H), 5.12 (t, J=10.0 Hz, 1H). ¹³C-NMR (125 MHz,CD₃OD): δ 11.54, 11.95, 15.27, 15.95, 16.15, 16.25, 16.30, 17.80, 22.32,22.46, 25.95, 26.01, 27.27, 27.44, 27.46, 27.80, 30.12, 30.19, 30.90,33.38, 33.66, 37.97, 40.78, 40.89, 52.91, 53.03, 59.29, 59.67, 121.39,121.41, 125.13, 125.14, 125.46, 126.15, 132.12, 136.27, 140.56, 140.64,171.70, 171.73, 173.64, 173.72, 173.88, 174.39; ES-MS: mass calcd forChemical Formula: C₂₆H₄₆N₄O₃S 494.73. Found (M+) m/z 495.3.

Example 48

Synthesis of((R)-2-(3-(3-ethoxy-3-oxopropyl)thioureido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-73)

In a 100 mL round bottom flask, to a suspension of ethyl-3-isothiocynatopropionate (159 mg, 1 mmol) and S-trans, trans-farnesyl-L-cysteine (325mg, 1 mmol) in THF (5 mL) was added N,N-diisopropyl-ethyl-amine (0.87mL, 5 mmol) dropwise. The solution was stirred at room temperatureovernight. The mixture was diluted with ethyl acetate (60 mL) and washedby 0.5 N HCl (10 mL×1), H₂O (10 mL×1) and brine (10 mL×1). The organiclayer was dried over Na₂SO₄ and concentrated in vacuo. The residue waspurified by preparative HPLC (220 mg, 45%) to yield Compound N-73.¹H-NMR (500 MHz, CD₃OD): δ 1.17 (t, J=5.0 Hz, 3H), 1.50 (s, 3H), 1.51(s, 3H), 1.57 (s, 3H), 1.59 (s, 3H), 1.86-2.06 (m, 8H), 2.54 (t, J=5.0Hz, 2H), 2.77 (dd, J=5.0, 15.0 Hz, 1H), 2.95-2.96 (m, 1H), 3.05-3.06 (m,1H), 3.16-3.20 (m, 1H), 3.68 (broad, 2H), 4.06 (q, J=5.0 Hz, 2H),5.00-5.01 (m, 2H), 5.09-5.14 (m, 2H). ¹³C-NMR (125 MHz, CD₃OD): δ 14.65,16.19, 16.31, 17.83, 25.97, 27.39, 27.80, 30.70, 33.22, 33.91, 34.80,40.80, 40.90, 57.89, 61.03, 61.71, 61.93, 121.35, 121.70, 125.16,132.11, 136.24, 140.52, 173.76, 174.47, 210.16; ES-MS: mass calcd forChemical Formula: C₂₄H₄₀N₂O₄S₂ 484.72. Found (M+) m/z 485.3, (M+Na) m/z507.3.

Example 49

Synthesis of((R)-2-(3-(2-carboxyethyl)thioureido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-74)

In a 100 mL round bottom flask, to a suspension of ethyl-3-isothiocynatopropionate (1 mmol) and S-trans, trans-farnesyl-L-cysteine (325 mg, 1mmol) in THF (5 mL) was added N,N-diisopropyl-ethyl-amine (0.87 mL, 5mmol) dropwise. The solution was stirred at room temperature overnight.The mixture was diluted with ethyl acetate (60 mL) and washed by 0.5 NHCl (10 mL×1), H₂O (10 mL×1) and brine (10 mL×1). The organic layer wasdried over Na₂SO₄ and concentrated in vacuo to afford crude CompoundN-74. The crude Compound N-74 obtained above was dissolved in THF (3 mL)and a solution of LiOH.H₂O (126 mg, 3 mmol) in H₂O (2 mL) was addedslowly at 0° C. The reaction was left for 4 h. The solution was thendiluted with ethyl acetate (60 mL) and washed by 0.5 N HCl (10 mL×1),H₂O (10 mL×1) and brine (10 mL×1). The organic layer was dried overNa₂SO₄ and concentrated in vacuo. The resulting residue was then furtherpurified by preparative HPLC (230 mg, 50%) to yield Compound N-74.¹H-NMR (500 MHz, CD₃OD): δ 1.50 (s, 3H), 1.51 (s, 3H), 1.57 (s, 3H),1.59 (s, 3H), 1.86-2.04 (m, 8H), 2.53 (t, J=6.0 Hz, 2H), 2.77 (dd,J=6.5, 14.0 Hz, 1H), 2.94-2.95 (m, 1H), 3.05-3.06 (m, 1H), 3.15-3.17 (m,1H), 3.67 (broad, 2H), 5.00-5.01 (m, 2H), 5.09-5.14 (m, 2H). ¹³C-NMR(125 MHz, CD₃OD): δ 16.17, 16.30, 17.81, 25.96, 27.39, 27.79, 30.68,31.57, 32.81, 33.20, 33.90, 34.56, 37.55, 40.80, 40.89, 57.93, 61.05,121.34, 121.70, 125.48, 132.11, 136.24, 140.52, 173.76, 174.55, 175.68;ES-MS: mass calcd for Chemical Formula: C₂₂H₃₆N₂O₄S₂ 456.66. Found (M+)m/z 457.2, (M+Na) m/z 479.2.

Example 50

Synthesis of((R)-2-(3-(carboxymethyl)ureido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-15)

In a 100 mL round bottom flask, to a suspension ofethyl-isocyanate-acetate (1 mmol) and S-trans, trans-L-cysteine (325 mg,1 mmol) in THF (5 mL) was added N,N-diisopropyl-ethyl-amine (0.87 mL, 5mmol) dropwise. The solution was stirred at room temperature overnight.The mixture was diluted with ethyl acetate (60 mL) and washed by 0.5 NHCl (10 mL×1), H₂O (10 mL×1) and brine (10 mL×1). The organic layer wasdried over Na₂SO₄ and concentrated in vacuo to afford crude CompoundN-15. The crude Compound N-15 obtained above was dissolved in THF (3 mL)and a solution of LiOH.H₂O (126 mg, 3 mmol) in H₂O (2 mL) was addedslowly at 0° C. The reaction was left for 4 h. The solution was thendiluted with ethyl acetate (60 mL) and washed by 0.5 N HCl (10 mL×1),H₂O (10 mL×1) and brine (10 mL×1). The organic layer was dried overNa₂SO₄ and concentrated in vacuo. The resulting residue was then furtherpurified by preparative HPLC (40 mg, 50%) to yield Compound N-15. ¹H-NMR(500 MHz, CD₃OD): δ 1.62 (bs, 6H), 1.68 (s, 3H), 1.70 (s, 3H), 1.99 (t,J=7 Hz, 2H), 2.06-2.17 (m, 6H), 2.82 (dd, J=7.0, 14.0 Hz, 1H), 2.95 (dd,J=5.0, 13.5 Hz, 1H), 3.18 (dd, J=7.0, 13.0 Hz, 1H), 3.28 (dd, J=5.0,12.0 Hz, 1H), 3.89 (bs, 2H), 4.50-4.54 (m, 1H), 5.10-5.15 (m, 2H), 5.24(t, J=7.0 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.1, 17.8, 25.9, 27.4,27.8, 30.6, 34.5, 40.8, 40.9, 42.5, 54.2, 121.7, 125.2, 125.5, 132.1,136.3, 140.5, 160.1, 174.1, 174.8; ES-MS: mass calcd for ChemicalFormula: C₂₁H₃₄N₂O₅S 426.6. Found (M+Na) m/z 449.3.

Example 51

Synthesis of((R)-2-(2-methoxy-2-oxoacetamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-10)

In a 100 mL round bottom flask, S-trans, trans-Farnesyl-L-cysteine (325mg, 1 mmol) and N,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixedin CH₂Cl₂ (10 mL). Methyl chloro oxoacetate (122 mg, 1 mmol) was addedto the reaction mixture. The reaction solution was stirred at roomtemperature overnight. Reaction solvent was removed by rotaryevaporation. The remained residue was dissolved in ethyl acetate (100mL) and washed with an NH₄Cl saturated solution (50 mL), dried overNa₂SO₄ and concentrated to afford a crude mixture. The crude mixture waspurified by preparative HPLC (66 mg, 15%) to yield Compound N-10. ¹H-NMR(500 MHz, CDCl₃): δ 1.60 (bs, 6H), 1.66 (s, 3H), 1.67 (s, 3H), 1.97 (t,J=7 Hz, 2H), 2.02-2.15 (m, 6H), 2.95 (dd, J=6.3, 14.2 Hz, 1H), 3.01 (dd,J=4.7, 14.2 Hz, 1H), 3.16-3.27 (m, 2H), 3.93 (bs, 3H), 4.82 (m, 1H),5.09 (m, 2H), 5.20 (t, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz, CDCl₃): δ 16.0,16.2, 17.7, 25.7, 26.4, 26.7, 30.0, 32.5, 39.6, 39.7, 52.0, 53.9, 119.2,123.7, 124.3, 131.4, 135.5, 140.6, 156.1, 160.2, 173.2; ES-MS: masscalcd for Chemical Formula: C₂₁H₃₃NO₅S 411.6. Found (M+Na) m/z 434.2.

Example 52

Synthesis of((R)-2-(2-ethoxy-2-oxoacetamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-13)

In a 100 mL round bottom flask, S-trans, trans-farnesyl-L-cysteine (325mg, 1 mmol) and N,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixedin CH₂Cl₂ (10 mL). Ethyl chloro oxoacetate (122 mg, 1 mmol) was added tothe reaction mixture. The reaction solution was stirred at roomtemperature overnight. Reaction solvent was removed by rotaryevaporation. The remained residue was dissolved in ethyl acetate (100mL) and washed with an NH₄Cl saturated solution (50 mL), dried overNa₂SO₄ and concentrated to afford a crude mixture. The crude mixture waspurified by preparative HPLC (66 mg, 15%) to yield Compound N-13. ¹H-NMR(500 MHz, CDCl₃): δ 1.40 (t, J=7.0 Hz, 3H), 1.60 (bs, 6H), 1.66 (s, 3H),1.68 (s, 3H), 1.97 (t, J=7 Hz, 2H), 2.02-2.15 (m, 6H), 2.95 (dd, J=6.5,14.0 Hz, 1H), 3.02 (dd, J=5.0, 16.0 Hz, 1H), 3.16-3.27 (m, 2H), 4.37 (q,J=7.0 Hz, 2H), 4.81 (m, 1H), 5.09 (m, 2H), 5.19 (t, J=7.5 Hz, 1H).¹³C-NMR (125 MHz, CDCl₃): δ 14.0, 16.0, 16.2, 17.7, 25.7, 26.4, 26.7,30.0, 32.5, 39.6, 39.7, 52.2, 63.6, 119.3, 123.7, 124.3, 131.3, 135.5,140.5, 156.5, 160.0, 174.1; ES-MS: mass calcd for Chemical Formula:C₂₂H₃₅NO₅S 425.2. Found (M+Na) m/z 448.2.

Example 53

Synthesis of((R)-2-(carboxyformamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-19)

In a 100 mL round bottom flask, S-trans, trans-Farnesyl-L-cysteine (325mg, 1 mmol) and N,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixedin CH₂Cl₂ (10 mL). Ethyl chloro oxoacetate (122 mg, 1 mmol) was added tothe reaction mixture. The reaction solution was stirred at roomtemperature overnight. Reaction solvent was removed by rotaryevaporation. The remained residue was dissolved in ethyl acetate (100mL) and washed with an NH₄Cl saturated solution (50 mL), dried overNa₂SO₄ and concentrated to afford a crude mixture. The crude reactionmixture and LiOH (126 mg, 3 mmol) were mixed in THF (3 mL) and water (3mL). The reaction solution was stirred at room temperature for 4 hours.Ethyl acetate (50 mL) was added and then washed with 1N HCl (20 mL×2)and brine (20 mL×2) sequentially. The ethyl acetate solution was driedby Na₂SO₄ and concentrated in vacuo to afford a crude mixture. The crudemixture was purified by HPLC (60 mg, 16%) to yield Compound N-19. ¹H-NMR(500 MHz, CD₃OD): δ 1.60 (bs, 6H), 1.68 (s, 3H), 1.73 (s, 3H), 1.99 (t,J=7 Hz, 2H), 2.02-2.15 (m, 6H), 2.88 (dd, J=8.5, 14.0 Hz, 1H), 3.08 (dd,J=4.0, 14.0 Hz, 1H), 3.15 (dd, J=5.5, 13.5 Hz, 1H), 3.28 (dd, J=5.5,13.0 Hz, 1H), 4.64 (dd, J=4.0, 7.5 Hz, 1H), 5.09-5.13 (m, 2H), 5.19 (t,J=7.5 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.1, 16.2, 26.0, 27.4, 27.8,30.2, 33.0, 40.8, 40.9, 53.7, 121.5, 125.1, 125.5, 132.1, 136.3, 140.7,160.3, 162.4, 173.0; ES-MS: mass calcd for Chemical Formula: C₂₀H₃₁NO₅S397.5. Found (M+Na) m/z 420.2.

Example 54

Synthesis of(1-[1-Carboxy-2-(3,7,11-trimethyl-dodeca-2,6,10-trienylsulfanyl)-ethylcarbamoyl]-cyclopropanecarboxylicacid methyl ester) (Compound N-52)

In a 100 mL round bottom flask, 1,1-Cyclopropanedicarboxylic acidmonomethyl ester (158 mg, 1.1 mmol),2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (1.1 mg, 1.1 mmol) and N,N-diisopropyl-ethyl-amine(0.52 mL, 3 mmol) were mixed in THF (5 mL). The reaction solution wasstirred at room temperature for ten minutes. S-trans,trans-Farnesyl-L-cysteine (325 mg, 1 mmol) was added to reactionmixture. The reaction solution was stirred at room temperatureovernight. Ethyl acetate (50 mL) was added and then washed withsaturated ammonium chloride aqueous solution (20 mL×2), DI water (20mL×2) and brine (20 mL×2) sequentially. The ethyl acetate solution wasdried by Na₂SO₄ and concentrated in vacuo to afford crude Compound N-52.The crude Compound N-52 was purified by HPLC (120 mg, 27%) to yieldCompound N-52. ¹H-NMR (500 MHz, MeOH-d₄): δ 1.45-1.48 (m, 4H), 1.50 (s,6H), 1.57 (s, 3H), 1.58 (s, 3H), 1.85-1.89 (m, 2H), 1.93-1.98 (m, 4H),2.01-2.05 (m, 2H), 2.78-2.91 (m, 2H), 3.06-3.15 (m, 2H), 3.62 (s, 3H),4.56 (t, J=5.0 Hz, 1H), 4.97-5.02 (m, 2H), 5.12 (t, J=7.5 Hz, 1H).¹³C-NMR (125 MHz, MeOH-d₄): δ 9.2, 16.14, 16.20, 17.80, 20.05, 25.95,27.31, 27.37, 27.78, 30.61, 33.62, 40.74, 40.88, 52.96, 53.97, 121.61,125.11, 125.45, 132.11, 136.28, 140.59, 170.81, 173.75, 174.39; ES-MS:mass calcd for Chemical Formula: C₂₄H₃₇NO₅S 451.2. Found (M+Na) m/z474.2.

Example 55

Synthesis of(1-[1-Carboxy-2-(3,7,11-trimethyl-dodeca-2,6,10-trienylsulfanyl)-ethylcarbamoyl]-cyclopropanecarboxylicacid) (Compound N-45)

In a 100 mL round bottom flask, the crude Compound N-52 (1 mmol) ofExample 54, and LiOH (126 mg, 3 mmol) were mixed in THF (3 mL) and water(3 mL). The reaction solution was stirred at room temperature for 4hours. Ethyl acetate (50 mL) was added and then washed with 1N HCl (20mL×2) and brine (20 mL×2) sequentially. The ethyl acetate solution wasdried by Na₂SO₄ and concentrated in vacuo to afford a crude mixture. Thecrude mixture was purified by HPLC (200 mg, 46%) to yield Compound N-45.¹H-NMR (500 MHz, MeOH-d₄): δ 1.47 (m, 4H), 1.50 (s, 6H), 1.57 (s, 6H),1.85-1.89 (m, 2H), 1.91-2.06 (m, 6H), 2.78-2.90 (m, 2H), 3.08-3.16 (m,2H), 3.25 (s, 1H), 4.57 (t, J=5.5 Hz, 1H), 4.97-5.02 (m, 2H), 5.12 (t,J=7.5 Hz, 1H). ¹³C-NMR (125 MHz, MeOH-d₄): δ 9.2, 16.16, 16.21, 17.81,20.40, 25.96, 26.59, 27.38, 27.78, 30.66, 33.58, 40.75, 40.88, 53.94,121.63, 125.12, 125.46, 132.11, 136.28, 140.60, 171.59, 173.70, 175.97;ES-MS: mass calcd for Chemical Formula: C₂₃H₃₅NO₅S 437.2. Found (M+Na)m/z 460.2.

Example 56

Synthesis of(2-[(1-Hydrazinocarbonyl-cyclopropanecarbonyl)-amino]-3-(3,7,11-trimethyl-dodeca-2,6,10-trienylsulfanyl)-propionicacid) (Compound N-75)

In a 100 mL round bottom flask, the crude Compound N-52 (1 mmol) ofExample 54, was added to 1M NH₂NH₂ in THF (10 mL, 10 mmol). The reactionsolution was stirred at room temperature overnight. The THF solution wasconcentrated in vacuo to afford a crude mixture. The crude mixture waspurified by HPLC (65 mg, 52%) to yield Compound N-75. ¹H-NMR (500 MHz,MeOH-d₄): δ 1.17-1.27 (m, 4H), 1.50 (s, 6H), 1.57 (s, 3H), 1.58 (s, 3H),1.85-1.88 (m, 2H), 1.91-2.03 (m, 6H), 2.70 (dd, J=9.0, 13.5 Hz, 1H),3.00 (dd, J=3.5, 13.5 Hz, 1H), 3.06-3.15 (m, 2H), 3.21 (s, 1H), 4.32(dd, J=3.5, 8.5 Hz, 1H), 4.97-5.02 (m, 2H), 5.14 (t, J=7.5 Hz, 1H).¹³C-NMR (125 MHz, MeOH-d₄): δ 9.2, 14.87, 15.72, 16.15, 16.28, 17.82,25.97, 27.49, 27.80, 30.16, 30.53, 34.90, 40.77, 40.89, 56.22, 121.67,125.21, 125.47, 132.09, 136.17, 140.11, 171.68, 172.35, 177.58; ES-MS:mass calcd for Chemical Formula: C₂₄H₃₇N₃O₄S 451.2. Found (M+Na) m/z474.2.

Example 57

Synthesis of(2-(3-Hydrazinocarbonyl-propionylamino)-3-(3,7,11-trimethyl-dodeca-2,6,10-trienylsulfanyl)-propionicacid) (Compound N-76)

In a 100 mL round bottom flask, the crude Compound N-39 (1 mmol) ofExample 43, was added to 1M NH₂NH₂ in THF (10 mL, 10 mmol). The reactionsolution was stirred at room temperature overnight. The THF solution wasconcentrated in vacuo to afford a crude mixture. The crude mixture waspurified by HPLC (60 mg, 56%) to yield Compound N-76. ¹H-NMR (500 MHz,MeOH-d₄): δ 1.50 (s, 6H), 1.57 (s, 3H), 1.58 (s, 3H), 1.85-1.88 (m, 2H),1.91-1.96 (m, 4H), 1.99-2.03 (m, 2H), 2.35 (t, J=7.5 Hz, 2H), 2.47 (t,J=7.5 Hz, 2H), 2.68 (dd, J=7.5, 13.5 Hz, 1H), 2.91 (dd, J=4.0, 13.5 Hz,1H), 3.09-3.12 (m, 2H), 4.33 (dd, J=4.5, 7.5 Hz, 1H), 4.97-5.00 (m, 2H),5.14 (t, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz, MeOH-d₄): δ 16.11, 16.26,17.79, 25.94, 27.50, 27.79, 30.69, 30.74, 32.60, 35.63, 40.78, 40.89,55.60, 121.80, 125.22, 125.47, 132.08, 136.14, 139.98, 173.73, 174.17,177.47; ES-MS: mass calcd for Chemical Formula: C₂₂H₃₇N₃O₄S 439.3. Found(M+Na) m/z 462.2.

Example 58

Synthesis of a mixture of(4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-3-methyl-4-oxobutanoicacid) (Compound N-22) and(4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2-methyl-4-oxobutanoicacid) (Compound N-21)

In a 100 mL round bottom flask, to a solution of S-trans,trans-farnesyl-L-cysteine (325 mg, 1 mmol) andN-methyl-succinic-anhydride (1 mmol) in CH₂Cl₂ (10 mL) was addedN,N-diisopropyl-ethyl-amine (0.87 mL, 5 mmol). The solution was stirredat room temperature for 2 h. The reaction was quenched by 1 N HCl (10mL) and the pH was adjusted to ˜2.0-3.0. The mixture was extracted withethyl acetate (15 mL×3). The organic layer was dried over Na₂SO₄ andconcentrated in vacuo. The residue was further purified by preparativeHPLC yielding a 6:4 mixture of regioisomeric compounds, N-22 (the3-methyl isomer) and N-21 (the 2-methyl isomer), wherein eachregioisomer is a 1:1 ratio mixture of R-R and S-R isomers, similar tothe regioisomeric mixture of compounds N-34 and N-33 in Example 41 (296mg, 67% yield). ¹H-NMR (500 MHz, CDCl₃): δ 1.14-1.24 (m, 3H), 1.53 (s,6H), 1.59 (s, 3H), 1.61 (s, 3H), 1.88-2.03 (m, 8H), 2.29-2.66 (m, 2H),2.72-3.01 (m, 3H), 3.07-3.16 (m, 2H), 4.59 (dd, J=5.0, 10.0 Hz, 0.5H),4.69 (dd, J=5.0, 10.0 Hz, 0.5H), 5.01 (m, 2H), 5.13 (dd, J=5.0, 15.0 Hz,1H), 6.52 (m, 0.5H), 6.70 (m, 0.5H), 8.80 (broad, 2H). ¹³C-NMR (125 MHz,CDCl₃): δ 15.00, 15.12, 15.56, 15.82, 15.98, 16.51, 16.69, 16.84, 24.71,25.36, 25.39, 25.50, 25.67, 28.74, 28.79, 31.42, 31.47, 31.53, 31.72,35.04, 35.48, 35.61, 36.59, 36.98, 37.64, 38.40, 38.60, 38.67, 50.50,50.65, 50.95, 51.02, 117.90, 118.26, 118.29, 122.64, 122.67, 122.71,123.27, 130.30, 130.35, 134.34, 134.38, 139.26, 139.27, 139.29, 139.33,170.86, 171.02, 174.01, 174.84, 174.96, 175.23, 175.40, 175.76, 177.16,179.36, 180.65; ES-MS: mass calcd for Chemical Formula: C₂₃H₃₇NO₅S439.61. Found (M+) m/z 440.3, (M+Na) m/z 462.3.

Example 58a

Synthesis of a mixture of((S)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-3-methyl-4-oxobutanoicacid) and((R)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-3-methyl-4-oxobutanoicacid) (Compound N-22)

In a 100 mL round bottom flask, to a solution of S-trans,trans-farnesyl-L-cysteine (325 mg, 1 mmol) andN-methyl-succinic-anhydride (1 mmol) in CH₂Cl₂ (10 mL) was addedN,N-diisopropyl-ethyl-amine (0.87 mL, 5 mmol). The solution was stirredat room temperature for 2 h. The reaction was quenched by 1 N HCl (10mL) and the pH was adjusted to ˜2.0-3.0. The mixture was extracted withethyl acetate (15 mL×3). The organic layer was dried over Na₂SO₄ andconcentrated in vacuo. The residue was further purified by preparativeHPLC yielding the 6:4 regioisomeric mixture of Compound N-22 andCompound N-21 in Example 58. This mixture was further purified bypreparative HPLC to yield a 1:1 ratio mixture of R-R and S-R isomers ofCompound N-22, similar to Compound C racemate in Examples 5 and 5a (135mg, 31%). ¹H-NMR (500 MHz, CD₃OD): δ 1.10-1.12 (m, 3H), 1.50 (s, 6H),1.57 (s, 3H), 1.60 (s, 3H), 1.86-2.06 (m, 8H), 2.22-2.28 (m, 1H),2.55-2.63 (m, 2H), 2.76-2.81 (m, 1H), 2.87-2.92 (m, 1H), 3.02-3.06 (m,1H), 3.13-3.17 (m, 1H), 4.45-4.50 (m, 1H), 4.98-5.03 (m, 2H), 5.13 (t,J=7.5 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.15, 16.24, 17.22, 17.80,18.41, 25.96, 27.40, 27.79, 30.10, 33.56, 33.65, 37.33, 37.93, 38.57,40.02, 40.79, 40.89, 53.11, 53.28, 121.60, 125.14, 125.46, 132.12,136.27, 140.48, 140.50, 173.98, 174.59, 178.25, 179.26; ES-MS: masscalcd for Chemical Formula: C₂₃H₃₇NO₅S 439.61. Found (M+) m/z 440.3,(M+Na) m/z 462.2.

Example 59

Synthesis of((R)-2-(4-amino-4-oxobutanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-26)

In a 100 mL round bottom flask, to a solution of S-trans,trans-farnesyl-L-cysteine methyl ester (339 mg, 1 mmol),4-(4,6-dimethoxy-1,3,5-triazin-2yl)-4-methylmorpholinium chloride(DMTMM, 332 mg, 1.2 mmol) and 4-amino-4-oxobutanoic acid (140 mg, 1.2mmol) in CH₂Cl₂ (5 mL) was added N,N-diisopropyl-ethyl-amine (0.52 mL, 3mmol). The solution was stirred at room temperature overnight. Themixture was diluted with ethyl acetate (60 mL) and washed sequentiallywith an NH₄Cl saturated solution (10 mL×1), H₂O (10 mL×1) and brine (10mL×1). The organic layer was dried over Na₂SO₄ and concentrated invacuo. The resulting residue dissolved in MeOH (3 mL) was added 5 N NaOH(3 mL) at room temperature. The reaction was left at room temperaturefor 10 min and the pH of the solution was adjusted to 3.0. The mixturewas extracted by ethyl acetate (50 mL×1). The organic layer was driedover Na₂SO₄ and the solvent was removed in vacuo. The residue was thenfurther purified by preparative HPLC (237 mg, 56%) to yield CompoundN-26. ¹H-NMR (500 MHz, CD₃OD): δ 1.50 (s, 3H), 1.51 (s, 3H), 1.57 (s,3H), 1.60 (s, 3H), 1.86-2.04 (m, 8H), 2.40-2.48 (m, 4H), 2.63 (dd,J=5.0, 10.0 Hz, 1H), 2.89 (dd, J=5.0, 15.0 Hz, 1H), 3.02-3.06 (m, 1H),3.14-3.21 (m, 1H), 4.48 (dd, J=5.0, 10.0 Hz, 1H), 4.98-5.01 (m, 2H),5.13 (t, J=10.0 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.15, 16.24,17.81, 25.96, 27.41, 27.80, 30.19, 31.67, 31.97, 33.48, 40.80, 40.89,53.43, 121.61, 125.15, 125.47, 132.12, 136.28, 140.52, 174.10, 174.70,177.43; ES-MS: mass calcd for Chemical Formula: C₂₂H₃₆N₂O₄S 424.60.Found (M+) m/z 425.3.

Example 60

Synthesis of a mixture of(4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-3,3-dimethyl-4-oxobutanoicacid) (Compound N-27) and(4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2,2-dimethyl-4-oxobutanoicacid) (Compound N-28)

In a 100 mL round bottom flask, to a solution of S-trans,trans-farnesyl-L-cysteine (325 mg, 1 mmol) and 2,2-dimethylsuccinicanhydride (1 mmol) in CH₂Cl₂ (10 mL) was addedN,N-diisopropyl-ethyl-amine (0.87 mL, 5 mmol). The solution was stirredat room temperature for 2 h. Then, the reaction was quenched by 1N HCl(10 mL) and pH was adjusted to 2.0˜3.0. The mixtures were extracted byethyl acetate (15 mL×3). The organic layer was dried over Na₂SO₄ andconcentrated in vacuo. The residue was further purified by preparativeHPLC to yield a mixture of regioisomeric compounds Compound N-27 andCompound N-28, wherein the ratio of N-28 to N-27 is 7:3 (386 mg, 85%yield). ¹H-NMR (500 MHz, CD₃OD): δ 1.16-1.23 (m, 6H), 1.50 (s, 6H), 1.57(s, 3H), 1.60 (s, 3H), 1.86-2.06 (m, 8H), 2.46-2.49 (m, 2H), 2.61 (m,1H), 2.86 (m, 1H), 3.05-3.06 (m, 1H), 3.13-3.15 (m, 1H), 4.44-4.47 (m,1H), 4.99-5.01 (m, 2H), 5.13 (t, J=10.0 Hz, 1H). ¹³C-NMR (125 MHz,CD₃OD): δ 16.16, 16.26, 17.81, 25.80, 25.83, 25.89, 25.96, 27.41, 27.80,30.15, 30.72, 33.29, 33.45, 40.80, 40.90, 41.67, 41.89, 44.91, 46.19,53.19, 53.34, 121.59, 121.61, 125.15, 125.47, 132.12, 136.27, 140.48,140.53, 173.41, 174.92, 179.71, 181.16; ES-MS: mass calcd for ChemicalFormula: C₂₄H₃₉NO₅S 453.64. Found (M+) m/z 454.3, (M+Na) m/z 476.2.

Example 61

Synthesis of a mixture of ((2S,3S)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2,3-dimethyl-4-oxobutanoicacid) (Compound N-30a), ((2S,3R)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2,3-dimethyl-4-oxobutanoicacid) (Compound N-30b), ((2R,3R)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2,3-dimethyl-4-oxobutanoicacid) (Compound N-30c) and ((2R,3S)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2,3-dimethyl-4-oxobutanoicacid) (Compound N-30d)

In a 100 mL round bottom flask, to a solution of S-trans,trans-farnesyl-L-cysteine (325 mg, 1 mmol) and 2,3-dimethylsuccinicanhydride (1 mmol) in CH₂Cl₂ (10 mL) was addedN,N-diisopropyl-ethyl-amine (0.87 mL, 5 mmol). The solution was stirredat room temperature for 2 h. Then, the reaction was quenched by 1N HCl(10 mL) and pH was adjusted to 2.0˜3.0. The mixtures were extracted byethyl acetate (15 mL×3). The organic layer was dried over Na₂SO₄ andconcentrated in vacuo. The residue was further purified by preparativeHPLC to yield a mixture of Compound N-30a, Compound N-30b, CompoundN-30c and Compound N-30d, wherein the ratio of N-30a:N-30b:N-30c:N-30dis 1:1:1:1 (259 mg, 57%). ¹H-NMR (500 MHz, CD₃OD): δ 1.05-1.10 (m, 6H),1.50 (s, 3H), 1.51 (s, 3H), 1.57 (s, 3H), 1.60 (s, 3H), 1.86-2.06 (m,8H), 2.49-2.65 (m, 3H), 2.84-2.88 (m, 1H), 3.06 (m, 1H), 3.13-3.15 (m,1H), 4.43-4.48 (m, 1H), 4.99-5.02 (m, 2H), 5.11-5.14 (m, 1H). ¹³C-NMR(125 MHz, CD₃OD): δ 14.26, 14.36, 14.66, 14.92, 16.15, 16.25, 16.27,16.76, 17.02, 17.21, 17.81, 25.96, 27.41, 27.44, 27.80, 30.02, 30.09,30.16, 30.72, 33.34, 33.65, 40.80, 40.89, 43.22, 43.33, 43.58, 43.65,44.52, 44.72, 45.07, 53.06, 53.10, 53.30, 121.58, 121.61, 121.63,125.14, 125.18, 125.47, 132.12, 136.25, 140.48, 173.88, 174.10, 177.61,178.13, 178.21, 179.02, 179.07, 179.12; ES-MS: mass calcd for ChemicalFormula: C₂₄H₃₉NO₅S 453.64. Found (M+) m/z 454.2, (M+Na) m/z 476.2.

Example 62

Synthesis of a mixture of((S)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-3-methyl-4-oxobutanoicacid and(R)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-3-methyl-4-oxobutanoicacid) (Compound N-21)

In a 100 mL round bottom flask, to a solution of S-trans,trans-farnesyl-L-cysteine methyl ester (339 mg, 1 mmol),4-(4,6-dimethoxy-1,3,5-triazin-2yl)-4-methylmorpholinium chloride(DMTMM, 332 mg, 1.2 mmol) and (R)-4-methoxy-2-methyl-4-oxobutanoic acid(175 mg, 1.2 mmol) in CH₂Cl₂ (5 mL) was addedN,N-diisopropyl-ethyl-amine (0.52 mL, 3 mmol). The solution was stirredat room temperature for 4 h. The mixture was diluted with ethyl acetate(60 mL) and washed sequentially with an NH₄Cl saturated solution (10mL×1), H₂O (10 mL×1) and brine (10 mL×1). The organic layer was driedover Na₂SO₄ and concentrated in vacuo. The residue was purified by flashchromatography on silica gel with hexanes/ethyl acetate (3/1) as eluent.The product obtained above was dissolved in THF (4 mL) and a solution ofLiOH.H₂O (203 mg, 4.83 mmol) in H₂O (2 mL) was added slowly at 0° C. Thereaction was left from 0° C. to room temperature overnight. The solutionwas then diluted with ethyl acetate (60 mL) and washed with 0.5 N HCl(10 mL×1), H₂O (10 mL×1) and brine (15 mL×1). The organic layer wasdried over Na₂SO₄ and concentrated in vacuo. The residue was thenfurther purified by preparative HPLC to yield a 1:1 ratio mixture of R-Rand S-R isomers of Compound N-21, similar to Compound C racemate inExamples 5 and 5a (150 mg, 34%). ¹H-NMR (500 MHz, CD₃OD): δ 1.08-1.12(m, 3H), 1.53 (s, 6H), 1.57 (s, 3H), 1.60 (s, 3H), 1.86-2.05 (m, 8H),2.23-2.28 (m, 1H), 2.55-2.63 (m, 2H), 2.77-2.78 (m, 1H), 2.85-2.86 (m,1H), 3.05-3.06 (m, 1H), 3.14-3.18 (m, 1H), 4.44-4.47 (m, 1H), 4.99-5.01(m, 2H), 5.13 (t, J=10.0 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.14,16.24, 17.22, 17.80, 18.09, 25.95, 27.40, 27.42, 27.79, 30.14, 33.34,37.40, 37.83, 38.72, 39.95, 40.79, 40.89, 53.35, 53.43, 121.60, 122.62,125.15, 125.16, 125.46, 132.11, 136.25, 140.48, 140.51, 174.04, 174.08,175.52, 178.29, 179.26; ES-MS: mass calcd for Chemical Formula:C₂₃H₃₇NO₅S 439.61. Found (M+) m/z 440.3, (M+Na) m/z 462.2.

Example 63

Synthesis of a mixture of stereoisomers((1S,2R)-2-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylcarbamoyl)cyclohexanecarboxylicacid) (Compound N-51a) and((1R,2S)-2-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylcarbamoyl)cyclohexanecarboxylicacid) (Compound N-51b)

In a 100 mL round bottom flask, to a solution of S-trans,trans-farnesyl-L-cysteine (325 mg, 1 mmol) and hexa-hydro-phthalicanhydride (1 mmol) in CH₂Cl₂ (10 mL) was addedN,N-diisopropyl-ethyl-amine (0.87 mL, 5 mmol). The solution was stirredat room temperature for 2 h. Then, the reaction was quenched by 1N HCl(10 mL) and pH was adjusted to 2.0˜3.0. The mixtures were extracted byethyl acetate (15 mL×3). The organic layer was dried over Na₂SO₄ andconcentrated in vacuo. The residue was further purified by preparativeHPLC to yield a 7:3 mixture of Compound N-51a and Compound N-51b (352mg, 73%). ¹H-NMR (500 MHz, CDCl₃): δ 1.18-1.48 (m, 4H), 1.53 (s, 6H),1.59 (s, 3H), 1.61 (s, 3H), 1.71-1.88 (m, 4H), 1.88-2.00 (m, 8H),2.79-2.89 (m, 3H), 3.01-3.18 (m, 3H), 4.51-4.77 (m, 1H), 5.01-5.03 (m,2H), 5.13 (m, 1H), 6.39 (m, 0.5H), 6.52 (d, J=5.0 Hz, 0.5H). ¹³C-NMR(125 MHz, CDCl₃): δ 15.00, 15.11, 15.18, 16.68, 20.77, 20.78, 21.32,22.53, 22.94, 23.90, 24.71, 25.38, 25.52, 25.67, 25.69, 27.06, 27.72,27.94, 28.20, 28.58, 28.65, 28.68, 28.72, 31.45, 31.64, 38.60, 38.65,38.67, 38.70, 38.80, 40.83, 41.09, 41.27, 42.11, 43.21, 49.61, 50.42,50.63, 52.79, 118.05, 118.24, 118.29, 122.67, 122.70, 122.77, 123.26,123.30, 130.28, 130.33, 134.27, 134.34, 134.37, 139.11, 139.27, 139.44,171.39, 173.68, 173.85, 178.08, 178.12, 178.69; ES-MS: mass calcd forChemical Formula: C₂₆H₄₁NO₅S 479.67. Found (M+) m/z 480.4, (M+Na) m/z502.3.

Example 64

Synthesis of((R)-2-acrylamido-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-42)

In a 100 mL round bottom flask, 3-chloro-propionic acid (1.0 mmol),coupling reagent (520 mg of PBOP or 380 mg of HATU, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) were mixed in CH₂Cl₂ (10mL). The reaction mixture was stirred at room temperature for 30minutes. S-trans, trans-Farnesyl-L-cysteine (325 mg, 1 mmol) was addedto the reaction mixture. The reaction mixture was stirred at roomtemperature overnight. CH₂Cl₂ was removed by rotary evaporation. Theresulting residue was dissolved in ethyl acetate (50 mL). The organicsolution was washed with an NH₄Cl saturated solution (50 mL), dried overNa₂SO₄, and concentrated to afford a crude mixture. The crude mixturewas purified by preparative HPLC (120 mg, 32%) to yield Compound N-42.¹H-NMR (500 MHz, CD₃OD): δ 1.50 (s, 6H), 1.57 (s, 3H), 1.64 (s, 3H),1.85-1.88 (m, 2H), 1.95-2.06 (m, 6H), 2.67 (dd, J=9.0, 14.0 Hz, 1H),2.93 (dd, J=4.5, 14.0 Hz, 1H), 3.06 (dd, J=7.0, 13.0 Hz, 1H), 3.20 (dd,J=9.0, 14.0 Hz, 1H), 4.58 (dd, J=4.5, 8.5 Hz, 1H), 4.98-5.03 (m, 2H),5.13 (t, J=7.5 Hz, 1H), 5.61 (d, J=10.0 Hz, 1H), 6.17 (d, J=17.0 Hz,1H), 6.28 (dd, J=10.0, 17.0 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.16,16.25, 17.81, 25.96, 27.39, 27.79, 30.16, 33.36, 40.79, 40.89, 53.50,121.60, 125.14, 125.47, 127.47, 131.65, 132.10, 136.27, 140.55, 168.00,173.81; ES-MS: mass calcd for Chemical Formula: C₂₁H₃₃NO₃S 379.56. Found(M+Na) m/z 402.2.

Example 65

Synthesis of a mixture of((R)-2-((S)-2-acetamido-4-ureidobutanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid and(R)-2-((R)-2-acetamido-4-ureidobutanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-77)

In a 100 mL round bottom flask, Fmoc-(D,L)-citrulline-OH (1 mmol) wasmixed with HATU (380 mg, 1 mmol) and N,N-diisopropyl-ethyl-amine (650mg, 5 mmol) in DMF (10 mL). After stirring at ambient temperature for 30min S-trans, trans-farnesyl-L-cysteine methyl ester (340 mg, 1 mmol) wasadded and the reaction mixture is additionally stirred for 16 hrs. Thereaction was quenched by addition of piperidine (10 mL) and stirring for2 hrs. Then water (10 mL) was added to crush the desired product out ofthe mixture followed by filtration. The separated product,(2S)-2-[4-(carbamoylamino)-2-aminobutanamido]-3-{[(2E,6E)-3,7,11-trimethyldodeca-2,6,10-trien-1-yl]sulfanyl}propanoicacid (468 mg, 1 mmol) was dissolved in acetic anhydride (3 mL, excess)and reaction was stirred at RT for 2 hrs. Then the excess of aceticanhydride was removed on rotavap, the resulting product was re-suspendedin THF (5 mL) and LiOH (saturated aq. solution, 0.25 mL) was added andthe resulting mixture stirred for 4 hrs. The mixture was purified byHPLC to yield a 1:1 racemic mixture of R-R and S-R isomers of CompoundN-77, similar to Compound C racemate in Examples 5 and 5a (209 mg, 41%yield). ¹H-NMR (500 MHz, MeOH-d₄): δ 1.24-1.61 (m, 9H), 1.63 (br s, 2H),1.89 (s, 3H), 1.93 (s, 3H), 1.91-2.05 (m, 2H), 2.52-2.55 (m, 1H),2.81-2.83 (m, 1H), 2.98-3.19 (m, 8H), 4.32 (t, J=4.5 Hz, 1H), 4.46 (t,J=6.5 Hz, 1H), 5.11 (br s, 2H), 5.23 (br s, 1H). ¹³C-NMR (125 MHz,MeOH-d₄): δ 16.1, 16.2, 23.2, 26.0, 27.4, 27.8, 30.2, 30.3, 33.8, 40.2,40.3, 48.5, 54.4, 54.5, 121.7, 125.1, 125.5, 132.1, 136.3, 140.5, 162.4,173.3, 174.5; ES-MS: mass calcd for Chemical Formula: C₂₆H₄₄N₄O₅S 524.7.Found (M+) m/z 525.3.

Example 66

Synthesis of a mixture of((S)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2-(3-ethylureido)-4-oxobutanoicacid and(R)-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-2-(3-ethylureido)-4-oxobutanoicacid) (Compound N-78)

In a 100 mL round bottom flask, Fmoc-(D,L)-aspartic acid alpha-methylester (1 mmol) was mixed with HATU (380 mg, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) in DMF (10 mL). Afterstirring at ambient temperature for 30 min, S-trans,trans-farnesyl-L-cysteine methyl ester (340 mg, 1 mmol) was added andthe reaction mixture is additionally stirred for 16 hrs. The reactionwas quenched by addition of piperidine (10 mL) and stirring for 2 hrs.Then water (10 mL) was added to crush the desired product out of themixture followed by filtration. The separated product,3-{[(1R)-1-carboxy-2-{[(2E,6E)-3,7,11-trimethyldodeca-2,6,10-trien-1-yl]sulfanyl}ethyl]carbamoyl}-2-aminopropanoicacid (440 mg, 1 mmol) was dissolved in ethyl isocyanate (3 mL, excess)and reaction was stirred at RT for 2 hrs. Then the reaction mixture wasconcentrated on rotavap, the resulting product was re-suspended in THF(5 mL) and LiOH (saturated aq. solution, 0.25 mL) was added and theresulting mixture stirred for 4 hrs. The mixture was purified by HPLC(167 mg, 32% yield) to yield a 1:1 racemic mixture of R-R and S-Risomers of Compound N-78, similar to Compound C racemate in Examples 5and 5a. ¹H-NMR (500 MHz, MeOH-d₄): δ 0.94 (t, J=7.5 Hz, 3H), 1.61 (s,6H), 1.63 (s, 6H), 2.55-2.81 (m, 4H), 2.83-2.86 (m, 1H), 3.04 (q, J=7.5Hz, 2H), 3.14-3.20 (m, 2H), 4.46-4.49 (m, 2H), 5.11 (m, 2H), 5.23 (t,J=7.5, 1H). ¹³C-NMR (125 MHz, MeOH-d₄): δ 16.1, 16.2, 18.3, 26.2, 27.4,27.8, 30.5, 34.4, 34.5, 35.8, 38.9, 40.8, 40.9, 54.0, 55.3, 121.5,125.1, 125.5, 132.1, 136.3, 140.5, 160.4, 172.4, 174.3, 175.6; ES-MS:mass calcd for Chemical Formula: C₂₅H₄₁N₃O₆S 511.7. Found (M+) m/z512.3.

Example 67

Synthesis of((R)-2-acetamido-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid and(S)-2-acetamido-4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid) (Compound N-32)

In a 100 mL round bottom flask, Fmoc-(D,L)-aspartic acid alpha-methylester (1 mmol) was mixed with HATU (380 mg, 1 mmol) andN,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) in DMF (10 mL). Afterstirring at ambient temperature for 30 min, S-trans,trans-farnesyl-L-cysteine methyl ester (340 mg, 1 mmol) was added andthe reaction mixture is additionally stirred for 16 hrs. The reactionwas quenched by addition of piperidine (10 mL) and stirring for 2 hrs.Then water (10 mL) was added to crush the desired product out of themixture followed by filtration. The separated product,3-{[(1R)-1-carboxy-2-{[(2E,6E)-3,7,11-trimethyldodeca-2,6,10-trien-1-yl]sulfanyl}ethyl]carbamoyl}-2-aminopropanoicacid (440 mg, 1 mmol) was dissolved in acetic anhydride (3 mL, excess)and reaction was stirred at RT for 2 hrs. Then the excess of aceticanhydride was removed on rotavap, the resulting product was re-suspendedin THF (5 mL) and LiOH (saturated aq. solution, 0.25 mL) was added andthe resulting mixture stirred for 4 hrs. The mixture was purified byHPLC (322 mg, 67% yield) to yield a 1:1 racemic mixture of R-R and S-Risomers of Compound N-32, similar to Compound C racemate in Examples 5and 5a. ¹H-NMR (500 MHz, MeOH-d₄): δ 1.29-1.68 (m, 12H), 1.73 (s, 3H),1.89-1.93 (m, 4H), 2.52-2.55 (m, 4H), 2.81-2.83 (m, 1H), 2.98-3.19 (m,2H), 4.32 (s, 1H), 4.46 (s, 1H), 5.11 (br s, 2H), 5.23 (br s, 1H).¹³C-NMR (125 MHz, MeOH-d₄): δ 16.1, 16.3, 17.8, 22.6, 23.2, 26.0, 27.4,27.8, 30.2, 30.3, 33.8, 40.2, 40.3, 48.5, 52.4, 121.7, 125.1, 125.5,132.1, 136.3, 140.5, 162.4, 172.0, 173.2; ES-MS: mass calcd for ChemicalFormula: C₂₄H₃₈N₂O₆S 482.6. Found (M+) m/z 483.3.

Example 68

Synthesis of racemic mixture of(2-{[(tert-butoxy)carbonyl]amino}-3-{[(2E,6E)-3,7,11-trimethyldodeca-2,6,10-trien-1-yl]oxy}propanoicacid) (Compound N-54)

In a 100 mL round bottom flask, N-Boc-(D,L)-serine (410 mg, 2 mmol) wasmixed with DMF (anhydrous, 10 mL) and NaH (60% in mineral oil, 100 mg,excess) with vigorous steering at under stream of nitrogen and at roomtemperature. After 30 min the excessive foaming subsided and trans,trans-farnesyl bromide was added dropwise (284 mg, 1 mmol) over 10 min.The reaction solution was stirred at room temperature overnight thenquenched with ammonium chloride (aq. sat., 20 mL) and the product wasextracted with ethyl acetate (2×10 mL). The organic layer was dried overmagnesium sulfate, concentrated and re-suspended in ethanol (1 mL) toafford a crude mixture. The crude mixture was purified by preparativeHPLC (76 mg, 19%) to yield a 1:1 racemic mixture of R and S enantiomersof Compound N-54. ¹H-NMR (500 MHz, MeOH-d₄): δ 1.47 (s, 9H), 1.69 (s,6H), 1.83 (s, 6H), 2.01 (t, J=6.5 Hz, 2H), 2.07-2.16 (m, 6H), 3.67 (dd,J=7.0, 12.0 Hz, 1H), 5.11-5.14 (m, 2H), 5.23 (t, J=7.5, 1H). ¹³C-NMR(125 MHz, MeOH-d₄): δ 16.3, 16.7, 17.9, 26.1, 27.4, 27.8, 28.8, 40.8,40.9, 55.3, 68.5, 70.4, 80.7, 121.8, 125.5, 132.1, 136.3, 141.8, 157.9,173.9; ES-MS: mass calcd for Chemical Formula: C₂₃H₃₉NO₅ 409.6. Found(M+Na) m/z 432.3.

Example 69

Synthesis of((R)-2-(cyclopropanesulfonamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-79)

In a 100 mL round bottom flask, to a suspension of cyclopropanesulfonylchloride (169 mg, 1.2 mmol) and S-trans, trans-farnesyl-L-cysteine (325mg, 1 mmol) in THF (5 mL) was added N,N-diisopropyl-ethyl-amine (0.52mL, 3 mmol) dropwise. The solution was stirred at room temperatureovernight. The mixture was diluted with ethyl acetate (60 mL) and washedby 0.5 N HCl (10 mL×1), H₂O (10 mL×2) and brine (10 mL×1). The organiclayer was dried over Na₂SO₄ and concentrated in vacuo. The residue wasthen further purified by preparative HPLC (50 mg, 12%) to yield CompoundN-79. ¹H-NMR (500 MHz, CD₃OD): δ 0.86-0.89 (m, 2H), 0.93-0.96 (m, 2H),1.50 (s, 3H), 1.51 (s, 3H), 1.57 (s, 3H), 1.61 (s, 3H), 1.86-1.89 (m,2H), 1.95-1.98 (m, 4H), 2.00-2.04 (m, 2H), 2.43-2.46 (m, 1H), 2.66-2.68(m, 1H), 2.76-2.79 (m, 1H), 3.09-3.13 (m, 1H), 3.16-3.21 (m, 1H), 4.02(t, J=5.0 Hz, 1H), 4.99-5.01 (m, 2H), 5.15 (t, J=5.0 Hz, 1H). ¹³C-NMR(125 MHz, CD₃OD): δ 5.58, 6.28, 16.16, 16.30, 17.81, 25.96, 27.39,27.80, 30.41, 31.73, 34.88, 40.79, 40.90, 57.75, 121.67, 125.17, 125.47,132.12, 136.28, 140.55, 174.39; ES-MS: mass calcd for Chemical Formula:C₂₁H₃₅NO₄S₂ 429.64. Found (M+) m/z 430.2, (M+Na) m/z 452.2.

Example 70

Synthesis of((R)-2-(2-carboxyethylsulfonamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-80)

In a 100 mL round bottom flask, to a solution of methyl3-(chlorosulfonyl)propanoate (187 mg, 1 mmol) and S-trans,trans-farnesyl-L-cysteine methyl ester (339 mg, 1 mmol) in THF (5 mL)was added N,N-diisopropyl-ethyl-amine (0.52 mL, 3 mmol) dropwise. Thesolution was stirred at 0° C. for 30 min and then room temperatureovernight. The mixture was diluted with ethyl acetate (60 mL) and washedby 0.5 N HCl (10 mL×1), H₂O (10 mL×1) and brine (10 mL×1). The organiclayer was dried over Na₂SO₄ and concentrated in vacuo. The resultingresidue was dissolved in THF (3 mL) and a solution of LiOH.H₂O (420 mg,10 mmol) in H₂O (2 mL) was added slowly at 0° C. The reaction was leftat room temperature overnight. The solution was then diluted with ethylacetate and washed by 0.5 N HCl (10 mL×1), H₂O (10 mL×2) and brine (15mL×1). The organic layer was dried over Na₂SO₄ and concentrated invacuo. The residue was then further purified by preparative HPLC (100mg, 22%) to yield Compound N-80. ¹H-NMR (500 MHz, CD₃OD): δ 1.50 (s,3H), 1.51 (s, 3H), 1.57 (s, 3H), 1.61 (s, 3H), 1.86-1.89 (m, 2H),1.95-2.00 (m, 4H), 2.01-2.06 (m, 4H), 2.65 (dd, J=8.0, 14.0 Hz, 1H),2.71-2.76 (m, 2H), 2.84 (dd, J=5.0, 14.0 Hz, 1H), 3.13 (dd, J=7.5, 13.5Hz, 1H), 3.24-3.29 (m, 1H), 4.05 (dd, J=5.0, 8.0 Hz, 1H), 4.98-5.03 (m,2H), 5.09-5.14 (t, J=8.0 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.15,16.30, 17.80, 25.95, 27.38, 27.79, 29.56, 30.44, 34.80, 40.78, 40.89,49.82, 57.51, 121.64, 125.16, 125.46, 132.12, 136.27, 140.62, 174.04,174.15; ES-MS: mass calcd for Chemical Formula: C₂₁H₃₅NO₆S₂ 461.64.Found (M+) m/z 462.2, (M+Na) m/z 484.2.

Example 71

Synthesis of(2-(N—((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethyl)sulfamoyl)benzoicacid) (Compound N-81)

In a 100 mL round bottom flask, to a solution of methyl2-(chlorosulfonyl)benzoate (281 mg, 1.2 mmol) and S-trans,trans-farnesyl-L-cysteine (325 mg, 1 mmol) in THF (5 mL) was addedN,N-diisopropyl-ethyl-amine (0.52 mL, 3 mmol) dropwise. The solution wasstirred at room temperature for 4 h. The mixture was diluted with ethylacetate (60 mL) and washed sequentially with an NH₄Cl saturated solution(10 mL×2), H₂O (10 mL×1) and brine (10 mL×1). The organic layer wasdried over Na₂SO₄ and concentrated in vacuo. The resulting residue wasdissolved in THF (3 mL) and a solution of LiOH.H₂O (210 mg, 5 mmol) inH₂O (2 mL) was added slowly at 0° C. The reaction was left at roomtemperature overnight. The reaction was quenched with 1 N HCl and pH wasadjusted to 2.0. The solution was then extracted by ethyl acetate (30mL×3). The organic layer was dried over Na₂SO₄ and concentrated invacuo. The residue was then further purified by preparative HPLC (290mg, 57%) to yield Compound N-81. ¹H-NMR (500 MHz, CD₃OD): δ 1.50 (s,6H), 1.55 (s, 3H), 1.57 (s, 3H), 1.86-1.89 (m, 2H), 1.94-2.02 (m, 6H),2.71 (dd, J=6.0, 14.0 Hz, 1H), 2.77 (dd, J=5.5, 14.0 Hz, 1H), 3.01-3.08(m, 2H), 4.10 (t, J=6.0 Hz, 1H), 5.00-5.06 (m, 3H), 7.56-7.62 (m, 2H),7.84 (d, J=6.5 Hz, 1H), 7.93 (d, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz,CD₃OD): δ 16.17, 16.28, 17.81, 24.24, 25.96, 27.36, 27.78, 30.52, 34.79,40.75, 40.89, 57.80, 121.53, 125.14, 125.46, 130.07, 132.13, 132.56,133.78, 136.28, 140.63, 140.72, 170.16, 173.07; ES-MS: mass calcd forChemical Formula: C₂₅H₃₅NO₆S₂ 509.68. Found (M+Na) m/z 532.1.

Example 72

Synthesis of(N-[1-Carboxy-2-(3,7,11,15-tetramethyl-hexadec-2-enylsulfanyl)-ethyl]-succinamicacid methyl ester) (Compound N-53)

In a 100 mL round bottom flask, phytol (trans: cis (2:1) isomericmixture of 34.9 mL, 100 mmol) and triethylamine (1.4 mL, 10 mmol) wereadded to toluene (100 mL), the reaction mixture was cooled down to −78°C. Phosphorus tribromide (4.7 mL, 50 mmol) was added dropwise. Afteraddition complete, the reaction mixture was warmed up to roomtemperature and stirred for 4 hours. Water (100 mL) was added dropwiseto quench the reaction. Ethyl acetate (200 mL) was added and then washedwith water (50 mL×2) and brine (50 mL×2) sequentially. The ethyl acetatesolution was dried by Na₂SO₄ and concentrated in vacuo. The resultingresidue was directly used for the next reaction. L-cysteinehydrochloride monohydrate (1.90 g, 10.73 mmol) and potassium carbonate(2.96 mg, 21.45 mmol) were added to ethanol (40 mL) and water (40 mL),the reaction was stirred at room temperature for 30 min, phytyl bromide(2.56 g, 7.15 mmol) was added. The reaction mixture was stirred at roomtemperature under argon for 4 hours. The precipitate obtained was washedby water, ethanol and dry in vacuum for 72 hours. White solid obtainedwas product which was directly used for the next reaction. Mono-methylsuccinate (132 mg, 1 mmol),2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (1.1 mg, 1.1 mmol) and N,N-diisopropyl-ethyl-amine(0.52 mL, 3 mmol) were mixed in THF (5 mL). The reaction solution wasstirred at room temperature for ten minutes.2-Amino-3-(3,7,11,15-tetramethyl-hexadec-2-enylsulfanyl)-propionic acid(399 mg, 1 mmol) was added to reaction mixture. The reaction solutionwas stirred at room temperature overnight. Ethyl acetate (50 mL) wasadded and then washed with saturated ammonium chloride aqueous solution(20 mL×2), DI water (20 mL×2) and brine (20 mL×2) sequentially. Theethyl acetate solution was dried by Na₂SO₄ and concentrated in vacuo toafford a crude mixture of 1:1 trans isomers and 1:1 cis isomers ofcompound N-53, wherein the ratio of trans isomers to cis isomers is 7:3(200 mg, 40%). ¹H-NMR (500 MHz, MeOH-d₄): δ 0.76-0.79 (m, 12H),1.00-1.46 (m, 19H), 1.58 and 1.63 (s, 3H), 1.91-1.99 (m, 2H), 2.48-2.52(m, 4H), 2.60-2.64 (m, 1H), 2.87 (dd, J=4.5, 14.0 Hz, 1H), 3.04-3.07 (m,1H), 3.14-3.18 (m, 1H), 3.57 (s, 3H), 4.46-4.49 (m, 1H), 5.12 (t, J=7.5Hz, 1H). ¹³C-NMR (125 MHz, MeOH-d₄): δ 20.12, 20.17, 20.23, 23.05,23.14, 23.59, 25.52, 25.94, 25.96, 26.30, 26.31, 26.64, 29.19, 30.20,30.40, 31.26, 32.87, 33.54, 33.79, 33.82, 33.88, 33.94, 33.97, 37.62,37.71, 38.41, 38.50, 40.56, 40.95, 52.27, 53.40, 53.53, 121.43, 121.89,140.87, 141.01; ES-MS: mass calcd for Chemical Formula: C₂₈H₅₁NO₅S513.3. Found (M+Na) m/z 536.3.

Example 73

Synthesis of(N-[1-Carboxy-2-(3,7,11,15-tetramethyl-hexadec-2-enylsulfanyl)-ethyl]-succinamicacid) (Compound N-48)

In a 100 mL round bottom flask, phytol (trans: cis (2:1) isomericmixture of 34.9 mL, 100 mmol) and triethylamine (1.4 mL, 10 mmol) wereadded to toluene (100 mL), the reaction mixture was cooled down to −78°C. Phosphorus tribromide (4.7 mL, 50 mmol) was added dropwise. Afteraddition complete, the reaction mixture was warmed up to roomtemperature and stirred for 4 hours. Water (100 mL) was added dropwiseto quench the reaction. Ethyl acetate (200 mL) was added and then washedwith water (50 mL×2) and brine (50 mL×2) sequentially. The ethyl acetatesolution was dried by Na₂SO₄ and concentrated in vacuo to afford a crudemixture of 1:1 trans isomers and 1:1 cis isomers of compound N-48,wherein the ratio of trans isomers to cis isomers is 7:3. The crudemixture (1 mmol) and LiOH (126 mg, 3 mmol) were mixed in THF (3 mL) andwater (3 mL). The reaction solution was stirred at room temperature for4 hours. Ethyl acetate (50 mL) was added and then washed with 1N HCl (20mL×2) and brine (20 mL×2) sequentially. The ethyl acetate solution wasdried by Na₂SO₄ and concentrated in vacuo to afford a partially purifiedmixture that was purified by HPLC to yield two fractions.

The first fraction yielded a mixture of 1:1 trans isomers and 1:1 cisisomers of compound N-48, wherein the ratio of trans isomers to cisisomers is 1:1 (50 mg, 20%). ¹H-NMR (500 MHz, MeOH-d₄): δ 0.76-0.79 (m,12H), 1.00-1.46 (m, 19H), 1.58 and 1.63 (s, 3H), 1.90-1.93 (m, 2H),2.46-2.49 (m, 4H), 2.62-2.66 (m, 1H), 2.87 (dd, J=4.5, 14.0 Hz, 1H),3.04-3.07 (m, 1H), 3.14-3.18 (m, 1H), 4.46-4.49 (m, 1H), 5.12 (t, J=7.5Hz, 1H). ¹³C-NMR (125 MHz, MeOH-d₄): δ 16.11, 20.13, 20.17, 20.23,23.06, 23.15, 23.59, 25.53, 25.95, 26.31, 26.65, 29.19, 30.25, 30.29,30.41, 31.41, 31.44, 32.88, 33.56, 33.79, 33.83, 33.94, 33.98, 37.62,37.72, 37.92, 38.01, 38.41, 38.47, 38.51, 40.57, 40.96, 53.43, 53.44,53.56, 121.44, 121.90, 140.86, 141.00, 174.02, 174.05, 174.47, 174.52,176.17, 176.19; ES-MS: mass calcd for Chemical Formula: C₂₇H₄₉NO₅S499.3. Found (M+Na) m/z 522.3.

The second fraction yielded a 1:1 mixture of trans isomers of compoundN-48 (45 mg, 23%). ¹H-NMR (500 MHz, MeOH-d₄): δ 0.76-0.79 (m, 12H),1.00-1.46 (m, 19H), 1.58 (s, 3H), 1.90-1.93 (m, 2H), 2.46-2.49 (m, 4H),2.63 (dd, J=8.5, 13.5 Hz, 1H), 2.87 (dd, J=4.5, 14.0 Hz, 1H), 3.02-3.07(m, 1H), 3.14-3.18 (m, 1H), 4.46-4.49 (m, 1H), 5.11 (t, J=7.5 Hz, 1H).¹³C-NMR (125 MHz, MeOH-d₄): δ 16.10, 16.11, 20.11, 20.16, 20.22, 23.05,23.14, 25.52, 25.94, 25.95, 26.30, 26.32, 29.19, 30.23, 30.23, 30.28,31.40, 33.54, 33.55, 33.79, 33.82, 33.94, 33.97, 37.61, 37.71, 38.40,38.47, 38.50, 38.53, 40.56, 40.95, 53.43, 121.44, 140.87, 174.03,174.53, 176.19; ES-MS: mass calcd for Chemical Formula: C₂₇H₄₉NO₅S499.3. Found (M+Na) m/z 522.3.

Example 74

Synthesis of((R)-2-(3-nitropropanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-97)

To a solution of 3-nitropropionic acid (143 mg, 1.2 mmol) and4-(4,6-dimethoxy-1,3,5-triazin-2yl)-4-methylmorpholinium chloride(DMTMM, 332 mg, 1.2 mmol) in CH₂Cl₂ (5 mL) was addedN,N-diisopropyl-ethyl-amine (0.52 mL, 3 mmol). After stirring for 10min, S-trans, trans-farnesyl-L-cysteine methyl ester (339 mg, 1.0 mmol)was added slowly. The solution was stirred at room temperature for 4 hand then diluted with ethyl acetate (60 mL). The solution was washedsequentially with an NH4Cl saturated solution (10 mL×1), H₂O (10 mL×1)and brine (10 mL×1). The organic layer was dried over Na₂SO₄ andconcentrated in vacuo. The residue was purified by flash chromatographyon silica gel with hexanes/ethyl acetate (3/1) as eluent. The product(408 mg, 0.93 mmol) obtained above was dissolved in THF (4 mL) and asolution of LiOH.H₂O (117 mg, 2.79 mmol) in H₂O (3 mL) was added slowlyat 0° C. The reaction was left at 0° C. for 30 min. The solution wasthen diluted with ethyl acetate (60 mL) and washed with 0.5 N HCl (10mL×1), H₂O (10 mL×2) and brine (10 mL×1). The organic layer was driedover Na₂SO₄ and concentrated in vacuo. The residue was then furtherpurified by preparative HPLC (288 mg, 68%) to yield Compound N-97.¹H-NMR (500 MHz, CD₃OD) δ 1.51 (s, 6H), 1.57 (s, 3H), 1.59 (s, 3H),1.86-1.89 (m, 2H), 1.96-2.04 (m, 6H), 2.64 (dd, J=8.5, 14.0 Hz, 1H),2.85-2.87 (m, 3H), 3.06-3.07 (m, 1H), 3.18 (dd, J=8.5, 13.5 Hz, 1H),4.50 (dd, J=5.0, 8.0 Hz, 1H), 4.63 (dd, J=5.0, 11.0 Hz, 2H), 4.99-5.01(m, 2H), 5.13 (t, J=8.0 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.14,16.22, 17.80, 25.95, 27.40, 27.80, 30.19, 32.80, 33.47, 40.79, 40.87,53.50, 71.03, 121.56, 125.14, 125.46, 132.12, 136.26, 140.57, 171.55,173.86; ES-MS: mass calcd for Chemical Formula: C₂₁H₃₄N₂O₅S 426.57.Found (M+1) m/z 427.3, (M+23) m/z 449.3.

Example 75

Synthesis of((R)-2-(3-(furan-2-yl)propanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-96)

To a solution of 3-(2-furyl) propionic acid (168 mg, 1.2 mmol) and4-(4,6-dimethoxy-1,3,5-triazin-2yl)-4-methylmorpholinium chloride(DMTMM, 332 mg, 1.2 mmol) in CH₂Cl₂ (5 mL) was addedN,N-diisopropyl-ethyl-amine (0.52 mL, 3 mmol). After stirring for 10min, S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was addedslowly. The solution was stirred at room temperature overnight and thendiluted with ethyl acetate (60 mL). The solution was washed by 0.5 N HCl(10 mL×1), H₂O (10 mL×1) and brine (10 mL×1). The organic layer wasdried over Na₂SO₄ and concentrated in vacuo. The residue was purified bypreparative HPLC (333 mg, 74%) to yield Compound N-96. ¹H-NMR (500 MHz,CD₃OD): δ 1.50 (s, 6H), 1.57 (s, 3H), 1.59 (s, 3H), 1.86-1.89 (m, 2H),1.95-2.06 (m, 6H), 2.51 (t, J=8.0 Hz, 2H), 2.57-2.62 (m, 1H), 2.83-2.869(m, 3H), 3.05 (dd, J=7.5, 13.5 Hz, 1H), 3.12-3.16 (m, 1H), 4.49 (dd,J=4.5, 8.0 Hz, 1H), 4.99-5.01 (m, 2H), 5.13 (t, J=7.5 Hz, 1H). ¹³C-NMR(125 MHz, CD₃OD): δ 16.15, 16.24, 17.80, 24.96, 25.95, 27.39, 27.79,30.11, 33.41, 35.08, 40.79, 40.89, 53.30, 106.28, 111.19, 121.61,125.14, 125.46, 132.12, 136.27, 140.49, 142.36, 155.72, 174.00, 174.78;ES-MS: mass calcd for Chemical Formula: C₂₅H₃₇NO₄S 447.63. Found (M+1)m/z 448.3, (M+23) m/z 470.2.

Example 76

Synthesis of((R)-2-(2-(5-hydroxy-1H-indol-3-yl)acetamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-39)

In 24 mL vial, 5-hydroxyl indole-3-acetic acid (191 mg, 1.0 mmol), HATU(380 mg, 1 mmol) and N,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) weremixed in THF (10 mL). The reaction mixture was stirred at roomtemperature for 30 minutes. S-trans, trans-Farnesyl-L-cysteine (325 mg,1 mmol) was added to the reaction mixture. The reaction mixture wasstirred at room temperature overnight. THF was removed by rotaryevaporation. The resulting residue was dissolved in ethyl acetate (50mL). The organic solution was washed with water (50 mL) and brain (50mL), dried over Na₂SO₄, and concentrated to afford a crude mixture. Thecrude mixture was purified by preparative HPLC (210 mg, 42%) to yieldCompound N-39. ¹H-NMR (500 MHz, CD₃OD): δ 1.49 (s, 9H), 1.56 (S, 3H),1.85-1.98 (m, 6H), 2.62 (dd, J=8.0, 14.0 Hz, 1H), 2.80 (dd, J=4.5, 14.0Hz, 1H), 2.88 (dd, J=7.0, 13.0 Hz, 1H), 2.98 (dd, J=8.5, 13.0 Hz, 1H),3.55 (dd, J=6.5, 22.5 Hz, 2H), 4.49 (dd, J=5.0, 8.0 Hz, 1H), 4.98 (bs,2H), 6.57 (d, J=8.5 Hz, 1H), 6.84 (s, 1H), 7.07 (d, J=6.5 Hz, 2H).¹³C-NMR (125 MHz, CD₃OD): δ 16.2, 17.8, 26.0, 27.3, 27.4, 27.8, 28.8,30.2, 33.3, 33.8, 40.7, 40.9, 53.4, 103.7, 108.2, 112.7, 112.8, 121.5,125.2, 125.5, 125.8, 129.3, 132.1, 133.0, 136.2, 140.5, 151.5, 173.9,174.9; ES-MS: mass calcd for Chemical Formula: C₂₈H₃₈N₂O₄S 498.3 (M+).Found (M+1) m/z 499.2.

Example 77

Synthesis of((R)-2-(3-(thiophen-2-yl)propanamido)-3-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)propanoicacid) (Compound N-31)

To a solution of 3-(2-thienyl) propanoic acid (187 mg, 1.2 mmol) and4-(4,6-dimethoxy-1,3,5-triazin-2yl)-4-methylmorpholinium chloride(DMTMM, 332 mg, 1.2 mmol) in CH₂Cl₂ (5 mL) was addedN,N-diisopropyl-ethyl-amine (0.52 mL, 3 mmol). After stirring for 5 min,S-trans, trans-farnesyl-L-cysteine (325 mg, 1 mmol) was added slowly.The solution was stirred at room temperature for 4 h and then dilutedwith ethyl acetate (60 mL). The solution was washed sequentially with anNH₄Cl saturated solution (15 mL×2), H₂O (10 mL×1) and brine (15 mL×1).The organic layer was dried over Na₂SO₄ and concentrated in vacuo. Theresidue was purified by preparative HPLC (310 mg, 67%) to yield CompoundN-31. ¹H-NMR (500 MHz, CD₃OD): δ 1.50 (s, 6H), 1.57 (s, 3H), 1.59 (s,3H), 1.86-1.89 (m, 2H), 1.96-2.06 (m, 6H), 2.54 (t, J=7.5 Hz, 2H),2.56-2.61 (m, 1H), 2.87 (dd, J=4.5, 14.0 Hz, 1H), 3.00-3.06 (m, 3H),3.11-3.15 (m, 1H), 4.49 (dd, J=5.0, 8.5 Hz, 1H), 5.00-5.01 (m, 2H), 5.12(t, J=7.5 Hz, 1H). ¹³C-NMR (125 MHz, CD₃OD): δ 16.16, 16.25, 17.81,25.96, 26.74, 27.40, 27.79, 30.13, 33.42, 38.74, 40.79, 40.89, 53.32,121.62, 124.36, 125.14, 125.47, 125.75, 127.81, 132.11, 136.27, 140.47,144.46, 174.98, 174.64; ES-MS: mass calcd for Chemical Formula:C₂₅H₃₇NO₃S₂ 463.70. Found (M+23) m/z 486.2.

Example 78

Synthesis of(2-[(2-aminophenyl)formamido]-3-{[(2E,6E)-3,7,11-trimethyldodeca-2,6,10-trien-1-yl]sulfanyl}propanoicacid) (Compound N-35)

Anthranilic acid (137 mg, 1 mmol) is mixed with HATU (380 mg, 1 mmol)and N,N-diisopropyl-ethyl-amine (650 mg, 5 mmol) in DMF (10 mL). Afterstirring at ambient temperature for 30 min, S-trans,trans-farnesyl-L-cysteine methyl ester (340 mg, 1 mmol) is added and thereaction mixture is additionally stirred for 16 hrs. Then LiOH(saturated aq. solution, 0.25 mL) was added and the resulting mixturestirred for 4 hrs. The mixture was purified by HPLC (107 mg, 24% yield)to yield Compound N-35. ¹H-NMR (500 MHz, MeOH-d₄): δ 1.42 (s, 6H), 1.54(s, 3H), 1.58 (s, 3H), 1.84-2.07 (m, 8H), 2.76 (dd, J=12.1 Hz, J=14.2Hz, 1H), 2.93 (s, J=7.4 Hz, J=14.2 Hz, 1H), 3.14 (d, J=12.1 Hz, 2H),4.32 (t, J=4.5 Hz, 1H), 4.51 (br.s, 2H), 4.59 (t, J=7.5 Hz, 1H),4.98-5.00 (m, 2H), 5.15 (t, J=12.1 Hz, 1H), 6.55 (t, J=7.4 Hz, 1H), 6.65(d, J=7.2 Hz, 1H), 7.10 (t, J=7.4 Hz, 1H), 7.41 (d, J=7.4 Hz, 1H).¹³C-NMR (125 MHz, MeOH-d₄): δ 16.3, 16.4, 23.3, 25.9, 26.0, 27.7, 27.8,30.4, 40.7, 40.9, 53.9, 117.1, 117.8, 118.3, 121.4, 125.1, 125.5, 129.3,132.1, 134.4, 136.2, 140.6, 150.4, 171.6, 175.9; ES-MS: mass calcd forChemical Formula: C₂₆H₄₄N₄O₅S 444.6. Found (M+Na) m/z 466.3.

Biological Examples

Described below are in vivo assays used to measure the biologicalactivity of provided compounds, including the anti-inflammatory orproinflammatory properties of the compounds, as measured by edemainhibition, erythema inhibition and MPO inhibition.

Example 79 Mouse Model of Inflammation-Edema, Erythema and MPOBackground

The mouse ear model of contact irritation has been established as anappropriate model to determine whether topically appliedanti-inflammatories inhibit the development of acute, chemically induceddermal irritation [see Van Arman, C. G. et al., Clin Pharmacol Ther,1974, 16: 900-4; Young et al., J Invest Dermatol, 1983, 80: 48-52;Tramposch et al., (Morgan D W, Marshall L A eds), Birkhäuser Verlag:Basel, 1999, pp 179-204; and Gordon et al., J Invest Dermatol, 2008,128: 643-54)]. Moreover, the mouse ear model has been used by variousgroups to identify and compare members of differing classes ofanti-inflammatory agents with multiple mechanisms of action (reviewed inTramposch et al., (Morgan D W, Marshall L A eds), Birkhäuser Verlag:Basel, 1999, pp 179-204). The commonly used end points of inflammationare edema (Young et al., J Invest Dermatol, 1983, 80: 48-52), (assayedby increase in ear thickness), neutrophil infiltration (which ismeasured by assaying for the neutrophil marker myeloperoxidase (“MPO”)(see Bradley et al., Blood, 1982, 60: 618-22) and erythema (skinredness). Using this mouse in vivo model for contact irritation, thepresent example demonstrates that certain isoprenyl compounds of thepresent invention, when topically applied exhibit in vivoanti-inflammatory or proinflammatory activities, as evidenced by theeffect on the commonly-used inflammatory end-points such as edema,erythema and neutrophil infiltration (MPO neutrophil marker) activities.The example can further be used to identify which structures possessphysical or chemical properties critical for inhibiting innateinflammation in the skin.

(a) Protocol—Edema Inhibition

The protocol for inducing in vivo acute contact inflammation on the earsof live mice has been described elsewhere (reviewed in Tramposch et al.,(Morgan D W, Marshall L A eds), Birkhäuser Verlag: Basel, 1999, pp179-204). In brief, mice were sedated and their ears were treated with1.2 μg/20 uL TPA (i.e., tetradecanoylphorbol-13-acetate). After 5minutes, we dosed these TPA-treated ears with a single 8 μg/20 uL dose,a 2 ug/20 uL dose, or both doses, of the isoprenyl compounds. After 24hours, the mice were sacrificed and edema was measured by takingmicrometer readings of each ear. The percent inhibition of edema wasdetermined by taking the average ear thickness of compound-treated earsand dividing it by the average thickness of 12 ears that only receivedTPA and subtracting that value from 100%. These values were correctedfor the thickness of normal, non TPA-treated mouse ears of littermatecontrols. Results demonstrating percent inhibition of edema forrepresentative compounds of the present invention are depicted inFIG. 1. ED₅₀ values were calculated as described in Gordon et al., JInvest Derm, 2008, 128: 643-654. ED₅₀ results for AFC and compound A aredepicted in FIG. 2.

(b) Protocol-Erythema Inhibition

Another well documented biomarker of skin inflammation is skin redness,termed erythema, which is caused by capillary congestion and dilation inresponse to various chemical and environmental insults (see Denig, N. I.et al., Postgrad Med, 1998; 103: 199-200, 207-8, 212-3). The protocolfor measuring erythema inhibition by isoprenyl compounds was developedin-house by utilizing the CR-400 chroma meter from Konica Minolta(http://www.konicaminolta.com/instruments/products/color/colorimeters/cr400-410/index.html).This instrument was used to measure the Δa* redness value from 6 mmbiopsy punches taken 24 hours post TPA/compound treatment as describedin the edema inhibition section above. The percent inhibition oferythema was determined by taking the average Δa* redness value ofcompound-treated ears and dividing it by the average Δa* value of 12ears that only received TPA and subtracting that value from 100%. Thesevalues were corrected for the Δa* value of non TPA-treated mouse ears oflittermate controls. Results demonstrating percent inhibition oferythema for representative compounds of the present invention aredepicted in FIG. 1. ED₅₀ values were calculated as described in Gordonet al., J Invest Derm, 2008, 128: 643-654. ED₅₀ results for AFC andcompound A are depicted in FIG. 2.

(c) Protocol-MPO Inhibition

To assay for inhibition of dermal neutrophil infiltration by isoprenylcompounds, a standard method was used (see Bradley et al., J InvestDermatol, 1982, 78: 206-209; Young et al., J Invest Dermatol, 1983, 80:48-52; De Young et al, Agents Actions, 1989, 26: 335-41; and Rao et al.,Inflammation, 1993, 17: 723-41). Briefly, we homogenized 6 mm biopsypunches taken from both compound-treated ears as well as TPA-treated andnon-treated control groups. We quantitated the levels of MPO by acolorimetric reaction that was measured spectrophotometrically. Thepercent inhibition of neutrophil infiltration by each isoprenyl compoundwas determined by comparing the average MPO levels in the presence andabsence of these compounds. The calculation for percent inhibition ofMPO was determined similar to that as described for calculating thepercent edema inhibition. Results demonstrating percent inhibition ofMPO for representative compounds of the present invention are depictedin FIG. 1. ED₅₀ values were calculated as described in Gordon et al., JInvest Derm, 2008, 128: 643-654. ED₅₀ results for AFC and compound A aredepicted in FIG. 2. Summary of activity ranges determined from an MPOactivity assay for compounds in Table 1 are presented in FIG. 3.

Described below are assays used to measure the biological activity ofprovided compounds, including the anti-inflammatory properties of thecompounds, as measured by inhibition of cytokine levels determined usinginflammatory models.

Example 80 TPA-Induced Mouse Ear Model of Inflammation Inhibition ofCytokine Levels

The protocol for inducing acute inflammation in mouse ears has beendescribed elsewhere (reviewed in Tramposch et al., (Morgan D W, MarshallL A eds), Birkhäuser Verlag: Basel, 1999, pp 179-204) and similar to theprotocol described in Example 79. Using this mouse in vivo model forcontact irritation, the present example demonstrates that certainisoprenyl compounds, when topically applied, exhibit in vivoanti-inflammatory activities, in part, by inhibiting the levels ofpro-inflammatory cytokines, such as TNF-α and IL-1β, resulting in theobserved effects on the inflammatory end-points of edema, erythema andneutrophil infiltration (MPO neutrophil marker) activities, asdemonstrated in Example 79. In brief, male Swiss Webster (ICR) mice10-12 weeks age (Hilltop Lab Animals) were used for these experiments (6animals per group). Mice received 1.2 μg/20 μl TPA dissolved in acetone[10 μl applied both to the dorsal and ventral surfaces of the mouse ear(20 μl total) using a solvent pipette] to each ear to induce acuteirritation. After 5 minutes, Compound A was applied at severalconcentrations in ethanol. After 24 hours treatment, mice wereeuthanized and 6-mm punch biopsy specimens were obtained from each ear,snap frozen in liquid nitrogen and stored in −80° C. until use. Earbiopsy specimens were homogenized with HTAB buffer using aBio-Pulverizer (MP Biomedicals, 2×45 sec at 4 m/s). Samples werecentrifuged at 10,000 rpm for 10 min at 4° C. Supernatants weresubjected to cytokine profiling by ELISA for the stimulated productionof TNF-α and IL-1β using protein standards for quantification. ED₅₀results (μg/ear) for TNF-α and IL-1β, obtained for Compound A using aTPA-induced mouse ear inflammation model are depicted in FIG. 4.

Example 81 LPS-TLR4-Induced Inflammation Model in HMEC-1 CellsInhibition of Cytokine Levels

The activation of Toll-like receptor 4 (TLR4) by lipopolysaccharide(LPS) induces the release of proinflammatory cytokines that arenecessary to mediate key immune and inflammatory responses (reviewed inYong-Chen et al., Cytokines, 2008, 42: 145-151). The present exampledemonstrates that certain isoprenyl compounds of the present inventioninhibit TLR4 inflammatory signaling pathways resulting in reduction ofproinflammatory cytokine release, for example of IL-8. HumanMicrovascular Endothelial cells (HMECs) were cultured in EC basal medium(EBM; Cambrex, Walkersville, Md.), supplemented with 0.5% fetal bovineserum (FBS), epidermal growth factor (EGF) (10 ng/mL) hydrocortisone (1μg/mL) and 100 U/mL penicillin/100 μg/mL streptomycin at 37° C. with 5%CO₂ (referred to as supplemented media). In order to avoid possibleimmunomodulating effects of these agents during agonist/antagonisttreatments, for some periods, cells were kept in EBM supplemented onlywith 0.5% FBS and penicillin/streptomycin without EGF or hydrocortisone(referred to as depleted media). Cells were plated at a concentration of0.25×10⁶ cells/well in supplemented media in 12-well plates. After cellswere allowed to adhere (6-8 hours), media was changed to depleted media.After 24 hours, depleted media was removed and fresh depleted mediacontaining various concentrations of Compound A in triplicate was addedto the appropriate wells. Two hours later, to induce a pro-inflammatoryresponse, LPS was added (100 μM) in separate wells (in triplicate)(Bender et al., Exp Dermatol, 2008, 17: 752-60; and Seiffert et al., JInvest Dermatol, 2006, 126: 1017-27). Cell cultures were examined forviability by Trypan blue exclusion and the reduction of3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS assay; Promega, Madison, Wis.) to determine the percentage ofviable cells of various treatment concentrations of Compound A. After 6hours of incubation, supernatants were harvested and assayed byenzyme-linked immunosorbent assays (ELISA) for the stimulated release ofIL-8 using appropriate protein standards (BD Pharmigen). IL-8 levels(pg/mL), obtained with Compound A using an LPS-TLR4-induced inflammationmodel in HMEC-1 cells are depicted in FIG. 5.

Example 82 ATPγS-Purinergic Receptor-Induced Inflammation Model inHMEC-1 Cells Inhibition of Cytokine Levels

ATP, serving as an extra-cellular signaling molecule, is known toactivate purinergic P2 receptors which are expressed on a variety ofcells involved in immune and inflammatory responses, including macro-and microvascular endothelial cells (ECs). During the pathophysiology ofinflammatory skin disorders, dermal microvascular ECs recruitinflammatory cells, including leukocytes, to the sites of inflammation,such as on the skin, triggered, in part, by the release ofproinflammatory mediators, such as IL-6 and MCP-1 (Swerlick et al., JInvest Dermatol, 1993, 100: 111S-115S). It has been previouslydemonstrated that the non-hydrolyzable analog of ATP, i.e., ATPγSinduces the production of proinflammatory cytokines in human dermalmicrocascular endothelial cells through the modulation of the P2purinergic receptor signaling (Seiffert et al., J Invest Dermatol, 2006,126: 1017-27). The protocol for inducing the production ofproinflammatory cytokines in human microvascular endothelial cells(HMECs) with ATPγS, as previously described, serves as a cell-basedmodel for studying the anti-inflammatory activities of test compounds.Using this cell-based model, the present example demonstrates thatcertain isoprenyl compounds of the present invention exhibitanti-inflammatory activity, as evidenced by the inhibition ofATPγS-induced-purinergic receptor-mediated release of proinflammatorymediators such as IL-8 and MCP-1. HMECs were cultured in EC basal medium(EBM; Cambrex, Walkersville, Md.), supplemented with 0.5% fetal bovineserum (FBS), epidermal growth factor (EGF) (10 ng/mL) hydrocortisone (1μg/mL) and 100 U/mL penicillin/100 μg/mL streptomycin at 37° C. with 5%CO₂ (referred to as supplemented media). In order to avoid possibleimmunomodulating effects of these agents during agonist/antagonisttreatments, for some periods, cells were kept in EBM supplemented onlywith 0.5% FBS and penicillin/streptomycin without EGF or hydrocortisone(referred to as depleted media). Cells were plated at a concentration of0.25×10⁶ cells/well in supplemented media in 12-well plates. After cellsare allowed to adhere (6-8 hours), media is changed to depleted media.After 24 hours, depleted media was removed and fresh depleted mediacontaining various concentrations of Compound A in triplicate was addedto the appropriate wells. Two hours later, to induce a pro-inflammatoryresponse, ATPγS was added (100 μM) in separate wells (in triplicate)(Bender et al., Exp Dermatol, 2008, 17: 752-60; and Seiffert et al., JInvest Dermatol, 2006, 126: 1017-27). Cell cultures were examined forviability by Trypan blue exclusion and the reduction of3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS assay; Promega, Madison, Wis.) to determine the percentage ofviable cells of various treatment concentrations of Compound A. After 6hours of incubation, supernatants were harvested and assayed byenzyme-linked immunosorbent assays (ELISA) for the stimulated release ofMCP-1, and IL-8 using appropriate protein standards (BD Pharmigen). IL-8levels (pg/mL), obtained with Compound A using an ATPγS-purinergicReceptor-induced Inflammation model in HMEC-1 cells are depicted in FIG.6. MCP-1 levels (pg/mL), obtained with Compound A using anATPγS-purinergic Receptor-induced Inflammation model in HMEC-1 cells aredepicted in FIG. 7.

Example 83 TPA-Induced Inflammation Model in NHEK Cells Inhibition ofCytokine Levels

The present example demonstrates that certain isoprenyl compounds of thepresent invention exhibit anti-inflammatory activity, as evidenced bythe inhibition of TPA-induced release of proinflammatory mediators suchas IL-8, in a human keratinocyte cell line (NHEK), similar to the effecton TPA-induced in vivo mouse ear model of inflammation as described inExample 80. NHEK cells were cultured in keratinocyte growth medium (KGM;Gibco, Carlsbad, Calif.), in a serum-free environment, supplemented withEGF (10 ng/mL), hydrocortisone (1 μg/mL), bovine insulin (5 μg/mL) andhuman pituitary gland extract (2 mL) at 37° C. with 5% CO₂. To avoid anypossible modulating effects of these agents during agonist/antagonisttreatments, cells were kept in KGM supplemented without EGF orhydrocortisone (depleted medium). Cells were plated at a concentrationof 0.25×10⁶ cells/mL in 12 well plates in supplemented media. After thecells were allowed to adhere (6-8 hours), media was changed to depletedmedia. After 24 hours, the depleted media was removed and fresh depletedmedia containing various concentrations of Compound A in triplicate wasadded to appropriate wells. After 8 hours, the media was changed tomedia without Compound A. After 16 hours, cell viability was determinedby Trypan blue exclusion and MTS assay to determine the percentviability of various treatment concentrations of Compound A. Cells werecultured in TPA (5 ng/mL) to induce a pro-inflammatory response andrelease of IL-8. After 5 hours of incubation, supernatants wereharvested and assayed by ELISA for the stimulated release of IL-8.Various concentrations of Compound A were added to tissue culture wellsin triplicate 2 hours before addition of TPA as well as cells notexposed to TPA. Cell viability was determined by Trypan blue exclusionand MTS assay 16 hours after stimulation in a duplicate experiment wherecells were washed and fresh media added without TPA or Compound A at theend of the stimulation period. IL-8 levels (pg/mL), obtained withCompound A using a TPA-induced inflammation model in NHEK cells aredepicted in FIG. 8.

Example 84 TNFα-Induced Inflammation Model in HUVEC Cells Inhibition ofTNFα-Induced Cytokine Release

TNF-α is a plieotropic cytokine with proinflammatory andimmunomodulatory functions. The pathogenic role of TNF-α in inflammationis mediated through the interaction of TNF-α with TNF receptors that inturn result in induction of proinflammatory cytokines, such as TNF-α(itself), IL-8 and others. The present example demonstrates that certainisoprenyl compounds of the present invention exhibit anti-inflammatoryactivity, as evidenced by the reduction of proinflammatory cytokinessuch as IL-8, mediated through TNF receptor mediated signaling in humanumbilical vein endothelial cells (HUVECs). HUVEC cells were cultured inendothelial growth medium-2 (EGM-2; Lonza; Walkersville, Md.), in a lowserum environment (2% FBS), and supplemented with EGM-2 Bullet Kit(Lonza) at 37° C. with 5% CO₂. To avoid any possible modulating effectsof these agents during agonist/antagonist treatments, cells were kept inEGM-2 supplemented without serum or growth factors (depleted medium).Cells were plated at a concentration of 1×10⁵ cells/mL in 96-well platesin supplemented media. After the cells were allowed to adhere (6-8hours), media will was changed to depleted media. Twenty-four hourslater, media was removed and fresh depleted media containing variousconcentrations of AFC, Compound A and Compound B in triplicate was addedto appropriate wells. After 30 minutes of pre-incubation, cells werestimulated with recombinant Human TNF-α (1×10⁴ U/mL; Millipore,Billerica, Mass.) to induce a pro-inflammatory response and release ofIL-8. After 4 hours of incubation, supernatants were harvested andassayed by ELISA for the stimulated release of IL-8. Cell viability wasdetermined by Trypan blue exclusion and MTS assay to determine thepercent viability of various treatment concentrations of AFC, Compound Aand Compound B. IL-8 levels (pg/mL), obtained with AFC, Compound A andCompound B using TNF-α-induced inflammation model in HUVEC cells aredepicted in FIG. 9.

Example 85 Effects on Ovalbumin-Challenged Flaky Tail Mouse Model forAtopic Dermatitis

The flaky tail mouse strain, carries a mutation in the gene for theepidermal protein filaggrin, comparable for the mutation underlyinghuman atopic dermatitis or eczema, and is, therefore, a model for thedisease (Fallon et al., Nat Genetics, 2009, 41: 602-608). Topicallychallenging these mice with ovalbumin results in a atopic dermatitislike condition, exhibiting eczema and increased skin levels of TH2 andthe cytokines IL4, IL5 and IL10, usually appearing 4-5 weeks followingovalbumin application. Using this model, the present exampledemonstrates the effectiveness of isoprenyl compounds of the presentinvention in inhibiting and/or reducing the various end-pointsassociated with atopic dermatitis. Exemplary end points include but arenot limited to skin flakiness, skin levels of TH2 and other cytokineslike IL4, IL5 and IL10. The protocol for cutaneous application ofOvalbumin to the intact skin of flaky tail mice has been describedelsewhere (Fallon et al., Nat Genetics, 2009, 41: 602-608). In brief,the abdomens of 3-5 week ovalbumin-challenged ft/ft mice (6 animals pergroups) are shaved 24 hours prior to cutaneous application andsuspensions of Ovalbumin (50 μg in 50 μl PBS) are applied to the abdomenaccording to a strict regimen as described previously (Fallon et al.,Nat Genetics, 2009, 41: 602-608). Two sets of experiments are conducted:in the first set, the mice are pretreated with the isoprenyl compoundsof the present invention prior to and during the application ofovalbumin to study the effects of preventing and inhibiting thedevelopment of AD phenotype; and in the second set, the mice are treatedwith the isoprenyl compounds following 4-5 weeks of ovalbumin treatmentwhen the phenotype appears to study the effects of the compounds intreating the symptoms. For each isoprenyl compound tested, the compoundis applied at several concentrations in ethanol to study dose dependenteffects. Following each experiment, mice are euthanized and 6-mm punchbiopsy specimens from each abdomen are harvested, snap frozen in liquidnitrogen and stored in −80° C. until use. The abdominal skin specimensare homogenized with HTAB buffer using a Bio-Pulverizer (MP Biomedicals,2×45 sec at 4 m/s). Samples are centrifuged at 10,000 rpm for 10 min at4° C. Supernatants are subjected to cytokine profiling by ELISA for thelevels of TH2, IL4, IL5, and IL10 using protein standards forquantification.

Described below are assays used to measure the biological activity ofprovided compounds, including the anti-psoriasis properties of thecompounds, as measured by inhibition of T-helper lymphocyte infiltrationdetermined using a psoriasis mouse model.

Example 86 K5.Stat3c Mouse Psoriasis Model Inhibition ofHelper-T-Lymphocyte Infiltration

The spontaneous and injury induced appearance of plaques having the fullpsoriatic phenotype in a transgenic mouse constitutively expressingsignal transducer and activator of transcription 3 (STAT3C) underregulation of the keratin-5 promoter in basal epidermal keratinocytes(“K5.Stat3c mice”) has been recently reported (Sano et al. (2005). NatMed 11(1): 43-9). In addition, the skin from these K5.Stat3c mice whenallografted to immunodeficient nude mice do not develop plaques unlessthey are co-engrafted with activated T-cells, as occurs when humanpsoriatic skin when grafted to Severe Combined Immunodeficiency (SCID)mice (Wrone-Smith et al., J Clin Invest, 1996, 98: 1878-1887; Nickoloffet al., Am J Pathol, 1999, 155: 145-158), establishing the necessaryinteraction between the altered epidermis and the immune system. TheseCD3+ T-helper cells play a critical role in the psoriatic pathogenesisby controlling infiltration of T-lymphocytes. The protocol for studyingthe CD3+ Helper-T expression using the K5.Stat3c psoriasis mouse modelhas been previously described (Sano et al., Nat Med, 2005, 11: 43-9).

Using this model, the present invention demonstrates that certainisoprenyl compounds of the present invention, when topically applied,exhibits efficacy in treating psoriasis, as evidenced by the inhibitionof T-helper cell infiltration in the K5.Stat3c psoriasis mouse model.Briefly, 5 mice per treatment group were used. Dorsal skin samples of7-9 wk-old K5.Stat3c mice were shaved 48 hours prior to tape stripping.Mice were then anesthetized with Avertin and received 30 strokes of tapestripping. Compound B, dexamethasone (Dex, positive control) or acetonevehicle control was topically applied to shaved area at indicated timesand doses as depicted in FIG. 10. Mice were injected with BrdU 30minutes prior to sacrifice at day 5, and skin sections collected forhistological assessment of dermal inflammatory infiltrates. The dosedependent inhibition in the number of CD3+ T-helper cells, obtained withCompound B using the K5.Stat3c psoriasis mouse model are depicted inFIG. 11.

Described below are assays used to measure the biological activity ofprovided compounds, including the anti-inflammatory properties of thecompounds, as measured by inhibition of ICMT.

Example 87 ICMT Inhibition

In the G-protein signaling pathways, for regulatory interactions tooccur, many of the signal transduction proteins, including virtually allG-proteins, first must be modified by the post-translational addition ofa C₁₅ farnesyl or a C₂₀ geranylgeranyl polyisoprenoid group in thioetherlinkage to a cysteine residue located at or near the carboxyl terminuswithin a so-called CAAX box or related cysteine-containing sequence.Carboxy-terminal polyisoprenoid cysteines that ultimately result fromthese modifications may be subject to methylesterification by a specificmembrane associated S-adenosylmethionine-dependent isoprenyl-S-isoprenylmethyltransferase (ICMT). Compounds that can inhibit these enzymaticreactions or otherwise alter the interactions among polyisoprenylatedsignal transduction proteins, such as G-proteins and the proteinregulatory targets with which they interact, or other intracellularsignaling proteins, may be used to mitigate leukocyte responses and,theoretically, to treat inflammatory-related conditions. (See e.g.,Volker, et al., Methods Enzymol, 1995, 250: 216-225).

The present example demonstrates that certain isoprenyl compounds of thepresent invention exhibit anti-inflammatory activity, as evidenced bythe inhibition of the enzymatic activity of ICMT thereby modulatingG-protein methylation.

Mouse brain extracts containing ICMT activity were prepared and %inhibition level of [³H]-AFC methylation was determined by the heptaneextraction method described previously (Volker et al., Methods, 1:283-287). Briefly, reaction mixture containing: 5 μl of protein (brainextract ˜40 μg), 2 μl AFC, 2 μl of IPC analog, 36 μl buffer A, and 5 μl[³H]-SAM (final concentration 10 μM) to a final volume of 50 μl, wasmixed and samples vortexed for 15 sec and then incubated for 30 min at37° C. Reaction was then quenched with 50 μl of 20% Tween20 (vortex for10 sec). Next 500 μl of heptane was added, the reaction mixture was thenvortexed for 10 sec and subsequently spun at 13,000 rpm for 5 min. Next,250 μl of the top layer was removed and placed in an open-top 1.5 μlcentrifuge tube. The open-top tubes were then spun for 30 min in avacuum centrifuge (Speed Vac Concentrator “Savant RH 4011”) to evaporateheptane. The tubes were then placed in 5 mL scintillation vials(containing 3 mL of scintillation fluid (Ecoscint, NationalDiagnostics). 200 μl of 1 M NaOH were added to each tube to hydrolyzethe base-labile AFCME and are immediately covered. The samples wereallowed to equilibrate overnight at 37° C., and then the levels of[³H]-MeOH that partitions into the cocktail, were quantified, by liquidscintillation spectrometry (Beckman LS 6500). Percent reduction of ICMTsubstrate, methylated acetyl-farnesyl-cysteine, obtained with compoundN-64, compound N-19, compound A, compound N-30 and compound N-77 aredepicted in FIG. 12.

Described below are assays used to measure the biological activity ofprovided compounds, including the anti-oxidant properties of thecompounds, as measured by inhibition of oxidative burst fromneutrophils, as determined by reduction of superoxide formation.

Example 88 Inhibition of Oxidative Burst from Neutrophils

Oxidative stresses caused by environmental insults such as ultraviolet(“UV”) rays from the sun, cigarette smoke exposure, consumption of foodswith high saturated fat and environmental pollutants as well as thenatural process of aging, contributing to the generation of freeradicals and reactive oxygen species (“ROS”), stimulate inflammatoryresponses, especially in the skin (Pilla et al., Intl J Cosm Sci, 2005,27: 17-34). High levels of ROS contribute to adverse effects on the skinincluding erythema, edema, photoaging and skin cancer (Trouba et al.Antioxid. Redox Signal 2002 v4 p 665-673). Neutrophil infiltrationduring inflammatory responses is associated with increased oxygenconsumption and generation of ROS. Extracellular inflammatory agonistssuch as fMLP bind to GPCRs such as formyl peptide receptors (“FPR”) totrigger the oxidative burst response (i.e., the rapid rapid release ofROS). Such oxidative burst responses from neutrophils are alsoassociated with irritable bowel syndrome, including ulcerative colitis(Keshavarzian et al., J Lab Clin Med, 1997, 130: 216-225).

The present invention demonstrates that certain isoprenyl compounds ofthe present invention exhibit anti-oxidant and anti-inflammatoryactivities, as evidenced by the inhibition of fMLP-induced GPCR-mediatedrelease of ROS.

The superoxide release assay is based on published protocols (Goldsteinet al., J Clin Invest, 1975, 56: 1155-63). Briefly, cells werepre-incubated for 10 min at 37° C. with a mixture of cytochrome c (75 μMfinal concentration), cytochalasin B (5 μg/mL) with or without SOD (10μg/mL) and with or without compounds (ranging from 0 to 100 μM). Toinitiate O₂ ⁻ release, fMLP (0.2 μM) was added the cells are incubatedfor 10 min at 37° C. Samples were then placed on ice for 5 min andsubsequently centrifuged at 3,000 rpm at 4° C. The supernatant was thenanalyzed by spectrophotometric measurement at 550 and 556.5 nm. Percentreduction of superoxide formation, obtained with AFC, compound C,compound N-25, AFC-methyl ester (AFC-ME) and AFC-AcetoxylMethane(AFC-AM) are depicted in FIG. 13.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, that while the invention hereinhas been described with reference to particular embodiments, it is to beunderstood that these embodiments are merely illustrative of theprinciples and applications of the present invention. It is therefore tobe understood that numerous modifications may be made to theillustrative embodiments and that other arrangements may be devisedwithout departing from the spirit and scope of the present invention asdefined by the appended claims.

In the claims articles such as “a,” “an,” and “the” may mean one or morethan one unless indicated to the contrary or otherwise evident from thecontext. Claims or descriptions that include “or” between one or moremembers of a group are considered satisfied if one, more than one, orall of the group members are present in, employed in, or otherwiserelevant to a given product or process unless indicated to the contraryor otherwise evident from the context. The invention includesembodiments in which exactly one member of the group is present in,employed in, or otherwise relevant to a given product or process. Theinvention includes embodiments in which more than one, or all of thegroup members are present in, employed in, or otherwise relevant to agiven product or process. Furthermore, it is to be understood that theinvention encompasses all variations, combinations, and permutations inwhich one or more limitations, elements, clauses, descriptive terms,etc., from one or more of the listed claims is introduced into anotherclaim. For example, any claim that is dependent on another claim can bemodified to include one or more limitations found in any other claimthat is dependent on the same base claim.

Where elements are presented as lists, e.g., in Markush group format, itis to be understood that each subgroup of the elements is alsodisclosed, and any element(s) can be removed from the group. It shouldit be understood that, in general, where the invention, or aspects ofthe invention, is/are referred to as comprising particular elements,features, etc., certain embodiments of the invention or aspects of theinvention consist, or consist essentially of, such elements, features,etc. For purposes of simplicity those embodiments have not beenspecifically set forth in haec verba herein. It is noted that the term“comprising” is intended to be open and permits the inclusion ofadditional elements or steps.

Where ranges are given, endpoints are included. Furthermore, it is to beunderstood that unless otherwise indicated or otherwise evident from thecontext and understanding of one of ordinary skill in the art, valuesthat are expressed as ranges can assume any specific value or subrangewithin the stated ranges in different embodiments of the invention, tothe tenth of the unit of the lower limit of the range, unless thecontext clearly dictates otherwise.

In addition, it is to be understood that any particular embodiment ofthe present invention that falls within the prior art may be explicitlyexcluded from any one or more of the claims. Since such embodiments aredeemed to be known to one of ordinary skill in the art, they may beexcluded even if the exclusion is not set forth explicitly herein. Anyparticular embodiment of the compositions of the invention (e.g., anytargeting moiety, any disease, disorder, and/or condition, any linkingagent, any method of administration, any therapeutic application, etc.)can be excluded from any one or more claims, for any reason, whether ornot related to the existence of prior art.

Publications discussed above and throughout the text are provided solelyfor their disclosure prior to the filing date of the presentapplication. Nothing herein is to be construed as an admission that theinventors are not entitled to antedate such disclosure by virtue ofprior disclosure.

1.-36. (canceled)
 37. A pharmaceutically acceptable salt of the compound4-(1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid, having the formula:


38. A pharmaceutical composition, comprising the pharmaceuticallyacceptable salt according to claim 37 and one or more pharmaceuticallyacceptable carriers.
 39. The pharmaceutical composition according toclaim 38, wherein said pharmaceutically acceptable salt of4-(1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid comprises at least 75% of a pharmaceutically acceptable salt of4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid.
 40. The pharmaceutical composition according to claim 39, whereinsaid pharmaceutically acceptable salt of4-(1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid comprises at least 80% of a pharmaceutically acceptable salt of4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid.
 41. The pharmaceutical composition according to claim 40, whereinsaid pharmaceutically acceptable salt of4-(1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid comprises at least 85% of a pharmaceutically acceptable salt of4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid.
 42. The pharmaceutical composition according to claim 41, whereinsaid pharmaceutically acceptable salt of4-(1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid comprises at least 90% of a pharmaceutically acceptable salt of4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid.
 43. The pharmaceutical composition according to claim 42, whereinsaid pharmaceutically acceptable salt of4-(1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid comprises at least 95% of a pharmaceutically acceptable salt of4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid.
 44. The pharmaceutical composition according to claim 43, whereinsaid pharmaceutically acceptable salt of4-(1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid comprises at least 97.5% of a pharmaceutically acceptable salt of4-((R)-1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid.
 45. The pharmaceutical composition according to claim 38, whereinsaid pharmaceutical composition is in a form suitable for oral,parenteral, urethral, vaginal, nasal, topical, pulmonary, deep lung,inhalation, buccal, or sublingual administration to a subject.
 46. Thepharmaceutical composition according to claim 45, wherein saidpharmaceutical composition is in a form suitable for oral, parenteral,or topical administration to a subject.
 47. The pharmaceuticalcomposition according to claim 46, wherein said pharmaceuticalcomposition is in a form suitable for topical administration to asubject.
 48. The pharmaceutical composition according to claim 47,wherein said pharmaceutical composition is in the form of an ointment,cream, lotion, paste, gel, spray, aerosol, or oil.
 49. Thepharmaceutical composition according to claim 48, wherein saidpharmaceutical composition is in the form of an ointment, cream, lotion,paste, or gel.
 50. A method of treating a disease or condition in asubject, comprising administering to said subject a pharmaceuticallyacceptable salt of4-(1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid, wherein said disease or condition is associated with aninflammatory response in said subject.
 51. The method according to claim50, wherein said pharmaceutically acceptable salt of4-(1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid is an alkali or alkaline earth salt.
 52. The method according toclaim 51, wherein said pharmaceutically acceptable salt is selected fromthe group consisting of sodium, lithium, potassium, calcium andmagnesium salt.
 53. The method according to claim 52, wherein saidpharmaceutically acceptable salt is the sodium salt.
 54. The methodaccording to claim 50, wherein said pharmaceutically acceptable salt of4-(1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid is administered to said subject in the form of a pharmaceuticalcomposition.
 55. The method according to claim 54, wherein saidpharmaceutical composition is administered to said subject by oral,parenteral, urethral, vaginal, nasal, topical, pulmonary, deep lung,inhalation, buccal, or sublingual administration.
 56. The methodaccording to claim 55, wherein said pharmaceutical composition isadministered to said subject by oral, parenteral, or topicaladministration.
 57. The method according to claim 56, wherein saidpharmaceutical composition is administered to said subject by topicaladministration.
 58. The method according to claim 50, wherein saiddisease or condition is an inflammatory skin condition.
 59. The methodaccording to claim 58, wherein said inflammatory skin condition isselected from rosacea, psoriasis, atopic dermatitis, and seborrhicdermatitis.
 60. The method according to claim 54, wherein said diseaseor condition is an inflammatory skin condition.
 61. The method accordingto claim 60, wherein said inflammatory skin condition is selected fromrosacea, psoriasis, atopic dermatitis, and seborrhic dermatitis.
 62. Themethod according to claim 61, wherein said inflammatory skin conditionis rosacea.
 63. The method according to claim 61, wherein saidinflammatory skin condition is psoriasis.
 64. The method according toclaim 61, wherein said inflammatory skin condition is atopic dermatitis.65. The method according to claim 61, wherein said inflammatory skincondition is seborrhic dermatitis.
 66. The method according to claim 50,wherein said pharmaceutically acceptable salt of4-(1-carboxy-2-((2E,6E)-3,7,11-trimethyldodeca-2,6,10-trienylthio)ethylamino)-4-oxobutanoicacid is administered to said subject in an amount from about 0.001 mg toabout 100 mg per kilogram of subject weight per day.
 67. Apharmaceutically acceptable salt of the compound having the formula:


68. The pharmaceutically acceptable salt according to claim 67, whereinsaid pharmaceutically acceptable salt is an alkali or alkaline earthsalt.
 69. The pharmaceutically acceptable salt according to claim 68,wherein said salt is selected from the group consisting of sodium,lithium, potassium, calcium and magnesium salt.
 70. The pharmaceuticallyacceptable salt according to claim 69, wherein said salt is the sodiumsalt.
 71. A pharmaceutical composition, comprising the pharmaceuticallyacceptable salt according to claim 67 and one or more pharmaceuticallyacceptable carriers.
 72. The pharmaceutical composition according toclaim 71, wherein said pharmaceutical composition is in a form suitablefor oral, parenteral, urethral, vaginal, nasal, topical, pulmonary, deeplung, inhalation, buccal, or sublingual administration to a subject. 73.The pharmaceutical composition according to claim 72, wherein saidpharmaceutical composition is in a form suitable for oral, parenteral,or topical administration to a subject.
 74. The pharmaceuticalcomposition according to claim 73, wherein said pharmaceuticalcomposition is in a form suitable for topical administration to asubject.
 75. The pharmaceutical composition according to claim 74,wherein said pharmaceutical composition is in the form of an ointment,cream, lotion, paste, gel, spray, aerosol, or oil.
 76. Thepharmaceutical composition according to claim 75, wherein saidpharmaceutical composition is in the form of an ointment, cream, lotion,paste, or gel.
 77. A method of treating a disease or condition in asubject, comprising administering to said subject a pharmaceuticallyacceptable salt of a compound having the formula:

wherein said disease or condition is associated with an inflammatoryresponse in said subject.
 78. The method according to claim 77, whereinsaid pharmaceutically acceptable salt is an alkali or alkaline earthsalt.
 79. The method according to claim 78, wherein saidpharmaceutically acceptable salt is selected from the group consistingof sodium, lithium, potassium, calcium and magnesium salt.
 80. Themethod according to claim 79, wherein said pharmaceutically acceptablesalt is the sodium salt.
 81. The method according to claim 77, whereinsaid pharmaceutically acceptable salt is administered to said subject inthe form of a pharmaceutical composition.
 82. The method according toclaim 81, wherein said pharmaceutical composition is administered tosaid subject by oral, parenteral, urethral, vaginal, nasal, topical,pulmonary, deep lung, inhalation, buccal, or sublingual administration.83. The method according to claim 82, wherein said pharmaceuticalcomposition is administered to said subject by oral, parenteral, ortopical administration.
 84. The method according to claim 83, whereinsaid pharmaceutical composition is administered to said subject bytopical administration.
 85. The method according to claim 77, whereinsaid disease or condition is an inflammatory skin condition.
 86. Themethod according to claim 85, wherein said inflammatory skin conditionis selected from rosacea, psoriasis, atopic dermatitis, and seborrhicdermatitis.
 87. The method according to claim 81, wherein said diseaseor condition is an inflammatory skin condition.
 88. The method accordingto claim 87, wherein said inflammatory skin condition is selected fromrosacea, psoriasis, atopic dermatitis, and seborrhic dermatitis.
 89. Themethod according to claim 88, wherein said inflammatory skin conditionis rosacea.
 90. The method according to claim 88, wherein saidinflammatory skin condition is psoriasis.
 91. The method according toclaim 88, wherein said inflammatory skin condition is atopic dermatitis.92. The method according to claim 88, wherein said inflammatory skincondition is seborrhic dermatitis.
 93. The method according to claim 77,wherein said pharmaceutically acceptable salt is administered to saidsubject in an amount from about 0.001 mg to about 100 mg per kilogram ofsubject weight per day.